biotechrabbit Product Catalog

Transcription

1 biotechrabbit Product Catalog

2 About biotechrabbit Scientists around the world are working to leap ahead of diseases and epidemics and to fuel our lives with innovations. Aside from brilliant minds and relentless passion, the success of science depends on the quality of the materials used. biotechrabbit is determined to offer the best products and innovative solutions to those who lead the progress. We seek to serve our customers with the newest life science products and services. We care deeply about our relationships with our partners and customers. It is our flexibility and our genuine concern that enable us to offer highly customized solutions for their specific requirements. We want our partners and customers to leap and lead the progress of life science. Our team of highly engaged scientists, experienced managers and business developers is happy to be a part of the scientific community that is leading with key technologies and conducting advanced research. Our way of making business is to combine the passion and pure curiosity of excellent researchers with the agile spirit of true entrepreneurs. biotechrabbit leap and lead Contact Us biotechrabbit GmbH Neuendorfstr. 24a Hennigsdorf Germany info@biotechrabbit.com Office: Fax: Commercial Register: Neuruppin (HRB 9381 NP) Managing Director: Dr. Bernd Haase Principal Office: Hennigsdorf Tax Office: Oranienburg VAT No./USt-Id: DE Tax No.: We highly appreciate your contacting us personally, by telephone, fax, post or . It is our priority to get back to you as quickly as possible. Ordering: Tel: Fax: order@biotechrabbit.com OEM and Antibody Service Tel: Fax: oem@biotechrabbit.com Technical Support: Tel: Tel: Fax: support@biotechrabbit.com Sales: Tel: Fax: sales@biotechrabbit.com Collaboration and Partnership: Tel: partnership@biotechrabbit.com Career Opportunities: Tel : hr@biotechrabbit.com 2 Bulk and custom formulations available Please contact oem@biotechrabbit.com

3 Table of Contents OEM and Other Services 4 Antibody Services 5 Standard PCR Product Selection Guide 6 Taq DNA Polymerase, recombinant, 5 U/µl 7 Taq DNA Polymerase, convenient, 5 U/µl 7 PCR Master Mix, 2x 8 Green Taq DNA Polymerase, 5 U/µl 9 Green PCR Master Mix, 2x 9 Red Taq DNA Polymerase, 1 U/µl 10 Red PCR Master Mix, 2x 10 Hot Start PCR Product Selection Guide 11 Hot Start Taq DNA Polymerase, 5 U/µl 12 Hot Start PCR Master Mix, 2x 12 Red Hot Start DNA Polymerase, 5 U/µl 13 Red Hot Start PCR Master Mix, 2x 14 Green Hot Start PCR Master Mix, 2x 14 High Fidelity and Long Range PCR Product Selection Guide 15 Pfu DNA Polymerase, 2.5 U/µl 16 Pfu PCR Master Mix, 2x 16 Long And High Fidelity PCR Enzyme Mix, 2.5 U/µl 17 Long Range PCR Master Mix, 2x 17 Reverse Transcription and RT-PCR Product Selection Guide 18 One Step RT-PCR Kit 19 NEW RevertUP Reverse Transcriptase, 100 U/µl 19 MMuLV Reverse Transcriptase, 200 U/µl 20 RNAse Inhibitor, 40 U/µl 20 Quantitative Real-Time PCR Product Selection Guide 21 QPCR Green Master Mix, 2x Intercalating-dye-based qpcr mixes without reference dye 23 QPCR SyGreen Master Mix LRox and HRox, 2x Intercalating-dye-based qpcr mixes with ROX 24 QPCR Probe Master Mix, 2x Probe-based qpcr mixes without reference dye 25 QPCR Probe-R Master Mix LRox and HRox, 2x Probe-based qpcr mixes with ROX 26 NEW QRT-PCR SyGreen One Step Kits, LRox and HRox 27 NEW QRT-PCR Probe-R One Step Kits, LRox and HRox 28 Direct PCR/Crude Sample PCR 29 U/µl 29 dntp Mixes and Sets 30 Nucleic Acid Purification 32 NEW UPzol RNA Isolation Solution 33 NEW GenUP Total RNA Mini Kit 33 NEW GenUP Plasmid Mini Kit 34 NEW GenUP gdna Mini Kit 35 NEW GenUP Plant DNA Kit 35 NEW GenUP Bacterial gdna Mini Kit 36 NEW GenUP Virus RNA Kit 37 NEW GenUP Virus RNA/DNA Kit 38 NEW GenUP PCR Cleanup Kit 39 NEW GenUP PCR/Gel Cleanup Kit 39 NEW GenUP Gel Extraction Kit 40 DNA Electrophoresis 41 DNA Electrophoresis Ladders and Loading Dyes 42 Ultrapure Enzymes for Molecular Biology 43 phi 29 DNA Polymerase, 10 U/µl 44 U/µl 45 T4 DNA Ligase, Rapid, 600 U/µl 46 Uracil DNA Glycosylase, 2 U/µl 47 Protein Electrophoresis 48 Protein Electrophoresis Ladders 49 Protein Purification Media 50 His-Tagged Protein Purification Media 51 Immobilized Metal Affinity Chromatography Media (IMAC) 52 Ion-Exchange Chromatography Protein Purification Media (IEX) 53 Size-Exclusion Chromatography Protein Purification Media (SEC) 54 info@biotechrabbit.com 3

4 OEM and Other Services Most of biotechrabbit molecular biology and proteomics products are available in bulk or as custom formulations. biotechrabbit welcomes all your OEM and bulk requests: Tel: Fax: oem@biotechrabbit.com You will receive answers to your questions about the availability of OEM products within three working days. We are a team of dedicated scientists, experienced managers and busi ness developers headquartered in Germany. To be ahead of the newest technologies and achievements in science, we closely collaborate with scientists who are leading key technologies and conducting advanced research. As a reliable OEM partner we offer the following: Custom formulations and bulk production of most of the products from our portfolio A choice of enzymes for standard PCR, high-fidelity PCR, reverse transcription, cloning and other applications Glycerin-free enzymes and antibodies, ready for lyophilization Enzymes for diagnostics kit development Lyophilization service using special formulations Development projects - cloning of proteins, expression optimization, purification scheme development Custom vialing, packaging and labeling Flexibility and outstanding personal attitude Production process documentati on and quality certificates Continuous technical support customers ion is to create continuous and sustainable value and added benefit for our customers. We want to serve our With the newest and most advanced life science technologies and products With services to help co -develop new applications or diagnostic tests To achieve maximum performance in research and shortest time to results biotechrabbit leap and lead 4 Bulk and custom formulations available Please contact oem@biotechrabbit.com

5 Price Duration Workflow Antibody Services We offer an exceptional quality and high capacity antibody production service, which is provided by qualified specialist who have been working in the field for more than 10 years. biotechrabbit welcomes all antibody related requests : Tel: Fax: oem@biotechrabbit.com Our antibody production services include Monoclonal antibodies Choice of animals: mouse and rat Starting material mg of your protein (or peptide) or the protein sequence Development of hybridoma cells using the provided protein Generation of monoclonal antibodies against membrane proteins using a special vector and proprietary cell immunization and screening technology Binding and activity screening using ELISA, western blot, FACS or special activity tests Production of a test sample of up to 10 mg Large-scale production by fermentation for producing up to 1kg of antibody Hybridoma supernatants are ready for screening in 4 weeks Tests are completed in 4 weeks (depending on test ordered) When subcloning has been ordered, t he first and second subcloning of hybrodoma cells are completed in 4 6 weeks Production of monoclonal antibodies: 2 weeks for 10 mg scale 4 weeks for 200 mg scale Up to 8 weeks for larger amounts Delivery of monoclonal antibodies in PBS buffer or Tris buffer, according to Variable dependent on the size of the project, the production scale and on the panel of required tests Polyclonal antibodies Choice of animals: rabbit, guinea pig, goat, and sheep others on request Starting material approximately 1 mg protein Production of a polyclonal antiserum Antibody purification and labeling, when required Visit our homepage for additional services The serum or the affinity purified antibodies are delivered in 4 weeks after we receive the protein Alternative immunization strategies available on request Variable dependent on additional services Benefits for the customer State-of-the-art antibody production: fermentation performed by a team with 10 years of experience. Shortest project time and a larger variety of monoclonal antibodies from which to select : our special immunization requires much shorter time than conventional methods (17 days). Antibodies on demand: more than 1800 hybridomas have already been produced. Highest purity: due to the use of genetically engineered protein A/G (lacking the albumin -binding domain) and cultivation in serum-free or protein-free media. After one-step purification with affinity chromatography, bovine IgGs and serum albumins cannot be detected in our monoclonal antibodies. Remaining aggregates can be quantitatively removed with preparative size - exclusion chromatography. Endotoxin-free antibodies: cgmp-compliant documentation can be supplied for preclinical trials. Yields of up to 500 mg of antibody per liter of culture per day : with our high-cell-density (>2 x 107 cells per ml) fermentation that uses an efficient two liter reactor systems, gram quantities can be delivered within weeks. High antibody production capacity : from 3 mg to over a kilogram. Short turnaround times: due to high-capacity production facility. No waiting times for customers : the antibody production process starts the day after receiving the starting materials. We are always ready for your project start with only short notice. Test orders of 10 mg antibody available at low prices. Production process documentation and quality certificates. Continuous technical support, flexibility and personal attitude. biotechrabbit leap and lead info@biotechrabbit.com 5

6 Standard PCR Product Selection Guide Standard PCR typically has the following parameters: Target length is 1 3 kb GC content of the template is 40 60% Template is pure, abundant and unproblematic Very high-fidelity of amplification is not required Unmodified Taq DNA polymerase is the most common enzyme for standard PCR. For simplified reaction setup, PCR master mixes which contain all necessary PCR reagents are off ered. Colored, high-density PCR buffers are available for direct loading of PCR products onto gels. The enzyme formulations in colored storage buffer s allow easy visualization of enzyme-containing reaction mixtures and monitoring the reaction set up process. Prod uct Main featur e Main PCR applicati on Sim p lifi ed setup Opti mi zation freed om Direct ge l load ing Colore d enzyme mixt ures Fide lit y Maxi mu m prod uct len gth TA clon ing Taq DNA Polym erase, reco mbinant, 5 U / µl MgCl 2 suppl ied in a separate vial for maxim um flexib ili ty Standar d Typi cal for Taq 3 5 k b Taq DNA Polym erase, conveni ent, 5 U / µl Reactio n Buff er con tai ns op timal MgCl 2 co ncen tra tion Standar d Typi cal for Taq 3 5 k b PCR Mast er Mi x, 2x Premix ed PCR reag en ts; ju st add templa te, pri mer s a nd water. Routi ne, hi g h- throu g hpu t Typi cal for Taq 3 5 k b Green Ta q D NA Polym erase, 5 U/ µl Green R eactio n Buffer for d irect loadin g o nto gel after PCR Standar d wi th direct g el loadin g Typi cal for Taq 3 5 k b Green PC R Master Mix, 2 x Red Taq D NA Polym erase, 1 U/ µl Red PC R Master Mix, 2 x Premix ed PCR reag en ts; ju st add templa te, pri mer s a nd water, i ncludi ng Green R eactio n Buffer for d irect loadin g o nto gel after the PCR Red -color ed Taq DNA Pol ym erase Storag e Buff er for visual ization of mix es con tai nin g the en zyme Premix ed PCR reag en ts; ju st add templa te, pri mer s a nd water, i ncludi ng Red Buffer for visual ization of mix es con tai nin g the en zyme Routi ne, hi g h- throu g hpu t wi th direct g el loadin g Standar d wi th en zyme visual ization Routi ne, hi g h- throu g hpu t wi th en zyme visual ization Typi cal for Taq Typi cal for Taq Typi cal for Taq 3 5 k b 3 5 k b 3 5 k b 6 Bulk and custom formulations available Please contact oem@biotechrabbit.com

7 Taq DNA Polymerase, recombinant, 5 U/µl BR BR U 500 U Routine amplification up to 4 kb RT-PCR TA cloning Taq DN A Pol ymera se, r ecom bina nt, 5 U/µ l 1.8 ml 10x R eactio n Buffer 1.5 ml 50 mm Mg Cl 2 Taq DN A Pol ymera se, r ecom bina nt, 5 U/µ l 2 x 1.8 m l 10x R ea ction Buffer 1.5 ml 50 mm Mg Cl 2 For maximum flexibility, MgCl2 is supplied separately High product yields in a wide application range Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications Taq DNA Polymerase, 5 U/µl, in Storage Buffer: 20 mm Tris-HCl (ph 8.0 at 25 C), 30 mm KCl, 0.1 mm DTT, 0.5% Tween 20, 50% (v/v) glycerol 10x Reaction Buffer: 670 mm Tris-HCl (ph 8.8 at 25 C), 166 mm (NH 4) 2SO4, 4.5% Triton X-100, 2 mg/ml gelatin biotechrabbit Taq DNA Polymerase is a first-choice thermostable DNA polymerase for all routine PCR applications, ensuring high product yields from various templates for targets up to 4 kb in size. Taq DNA polymerase is a thermostable, highly processive 5' 3' DNA polymerase that has low 5 ' 3' exonuclease activity and lacks 3' 5' exonuclease (proofreading) activity. The latter allows incorporation of modified nucleotides into DNA. The enzyme also exhibits deoxynucleotidyl transferase activity, resulting in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhang s. UNIT DEFINITION One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dntp into acid - insoluble form in 30 minutes at 72 C in the presence of 1x Reaction Buffer. Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay Taq DNA Polymerase, convenient, 5 U/µl BR U Routine amplification up to 4 kb RT-PCR TA cloning Taq DN A Pol ymera se, co nv eni en t, 5 U /µl 2 x 1.8 m l 10x R ea ction Buffer wi th M g Cl 2 Optimal MgCl2 concentration in the Reaction Buffer High product yields in a wide application range Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications Taq DNA Polymerase, 5 U/µl, in Storage Buffer: 20 mm Tris-HCl, 100 mm NaCl, 1.0 mm DTT, 0.1 mm EDTA, Stabilizer, 50% glycerol, ph 7.5 at 25 C 10x Reaction Buffer with MgCl 2: 100 mm Tris-HCl (ph 8.3 at 25 C), 500 mm KCl, 15 mm MgCl 2 biotechrabbit Taq DNA Polymerase is a first -choice thermostable DNA polymerase for all routine PCR applications, ensuring high product yields from various templates for targets up to 4 kb in size. Taq DNA polymerase is a thermostable, highly processive 5' 3' DNA polymerase that has low 5 ' 3' exonuclease activity and lacks 3' 5' exonuclease (proofreading) activity. The latter allows incorporation of modified nucleotides into DNA. The enzyme also exhibits deoxynucleotidyl transferase activity, resulting in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs. Purified from a recombinant E. coli strain carrying the Taq DNA polymerase gene from Thermus aquaticus YT-1. UNIT DEFINITION One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dntp into acid - insoluble form in 30 minutes at 72 C in the presence of the 1x Reaction Buffer. Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rdna Contamination Test Functional PCR Assay info@biotechrabbit.com 7

8 PCR Master Mix, 2x BR BR rea ction s ( 50 µl) 500 r ea cti ons Routine amplification up to 4 kb TA cloning 2 x 1.25 ml PCR Master Mix, 2x 10 x 1.25 m l PCR Master Mix, 2x Exceptionally pure Taq DNA Polymerase and highest quality dntps and buffer in a 2x PCR Master Mix formulation High product yields and robustness in a wide application range PCR Master Mix, 2x: Proprietary composition contains all necessary PCR reagents, including 3 mm MgCl2. (1x has 1.5 mm MgCl2) biotechrabbit PCR Master Mix is a perfect choice for fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for routine, high-throughput PCR amplification of kb DNA targets. The 2x Master Mix contains highly purified recombinant biotechrabbit Taq DNA polymerase, highest quality dnt Ps and optimal PCR buffer reagents; thus, only template DNA, PCR primers and PCR-grade water need to be added. Taq DNA polymerase is a thermostable highly processive 5' 3' DNA polymerase that has low 5 ' 3' exonuclease activity and lacks 3 ' 5' exonuclease (proofreading) activity. The latter allows incorporation of modified nucleotides into DNA. The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay PCR Master Mix: Functional PCR Assay L Picture 1. Performance of biotechrabbit PCR Mas ter Mixes. A 700 bp genomic DNA fragment was amplified using different PCR master mixes. Colored PCR master mixes show comparable performance to colorless mixes. L 100 bp DNA Ladder, rtu (BR ) 1 PCR Master Mix, 2x (BR ) 2 Red PCR Master Mix, 2x (BR ) 3 Green PCR Master Mix, 2x (BR ) 4 Hot Start PCR Master Mix, 2x (BR ) 5 Red Hot Start PCR Master Mix, 2x (BR ) 6 Green Hot Start PCR Master Mix, 2x (BR ) Bulk and custom formulations available Please contact oem@biotechrabbit.com

9 Green Taq DNA Polymerase, 5 U/µl BR BR BR BR BR U 500 U 5 x 5 00 U 10 x 5 00 U 20 x 500 U 20 µl Taq DNA Pol ym erase, 5 U /µl 2 x 1.8 m l 5 x Gr een R ea ction Buffer 1.5 ml 50 mm Mg Cl µl Ta q DNA Pol ym erase, 5 U /µ l 4 x 1.8 ml 5x Gr een Rea ctio n Buffer 1.5 ml 50 mm Mg Cl 2 5 x 1 00 µl Taq DNA Pol ym erase, 5 U /µ l 5 x 1.8 ml 5x Gr een Rea ctio n Buffer 5 x 1.5 m l 50 mm Mg Cl2 10 x 1 00 µl Taq DNA Pol ym erase, 5 U /µl 10 x 1.8 m l 5x Gr een Rea ction Buffer 10 x 1.5 ml 5 0 mm MgC l 2 20 x 100 µl Taq DN A Pol ymerase, 5 U/µl 20 x 1.8 ml 5 x Green R eacti on Buff er 20 x 1.5 ml 5 0 mm M gcl 2 High-throughput PCR for immediate gel analysis Routine amplification up to 4 kb TA cloning Green Reaction Buffer allows direct loading on to a gel immediately after PCR High product yields and robustness Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications Taq DNA Polymerase, 5 U/µl, in Storage Buffer: 20 mm Tris-HCl (ph 8.0 at 25 C), 30 mm KCl, 0.1 mm DTT, 0.5% Tween 20, 50% (v/v) glycerol 5x Green Reaction Buffer wit h MgCl2: 335 mm Tris-HCl (ph 8.8 at 25 C), 83 mm (NH 4) 2SO4, 2.25% Triton -X-100, 1 mg/ml gelatin, t racking dyes (yellow and blue) biotechrabbit Green Taq DNA Polymerase is a first -choice thermostable DNA polymerase for routine PCR applications, allowing direct electrophoresis without the need to add loading buffer and ensuring high product yields from various templates. The 5x Green Taq Reaction Buffer is recommended for any amplification reaction that will be visualized by agarose gel electrophoresis and ethidium bromide staining. The Green Reaction Buffer contains two dyes (blue and yellow) that separate during electrophoresis, allowing the migration progress to be monitored. Reactions with Green Reaction Buffer have sufficient den sity for direct loading onto agarose gels. The dyes absorb between nm, making standard A 260 readings to determine DNA concentration unreliable. Taq DNA polymerase is a thermostable highly processive 5' 3' DNA polymerase that has low 5 ' 3' exonuclease activity and lacks 3' 5' exonuclease (proofreading) activity. The enzyme also exhibits deoxyribonucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs. UNIT DEFINITION One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dntp into acid - insoluble form in 30 minutes at 74 C in the presence of the reaction buffer. Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay Green PCR Master Mix, 2x BR BR rea ction s 500 r ea cti ons 2 x 1.25 ml Gr een PCR Master Mix, 2x 10 x 1.25 m l Green PCR Master Mix, 2x High-throughput PCR for immediate gel analysis Routine amplification up to 4 kb TA cloning Green PCR Master Mix formulation allows direct loading on to a gel immediately after PCR Exceptionally pure Taq DNA Polymerase and highest quality dntps in a 2x Green PCR Master Mix High product yields and robustness in a wide application range Green PCR Master Mix, 2x: Proprietary master mix composition contains all necessary PCR components, including 3 mm MgCl2 (1x has 1.5 mm MgCl 2) and blue and yellow electrophoresis dyes biotechrabbit Green PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the special formulation allows reactions onto be loaded onto the gel directly after amplification without a separate step for adding loading dye. PCR Master Mix contains two dyes (blue and yellow) that separate during electrophoresis, allowing the migration progress to be monitored. Reactions with the Green PCR Master Mix have sufficient density for direct loading on to agarose gels. Green Reaction Buffer also allows mixtures containing the enzyme to be identified. The Green PCR Master Mix contains highly purified recombinant biotechrabbit Taq DNA polymerase, highest quality dntps and optimal PCR buffer reagents; thus, only template DNA, PCR primers and PCR-grade water need to be added. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Green PCR Master Mix: Functional PCR Assay info@biotechrabbit.com 9

10 Red Taq DNA Polymerase, 1 U/µl BR BR BR BR BR U 500 U 5 x 500 U 10 x 500 U 20 x 500 U 100 µl R ed Taq DNA Pol ym erase, 1 U /µ l 1.8 ml 10x R eactio n Buffer 1.5 ml 50 mm Mg Cl µl Red Taq DNA Polymerase, 1 U /µl 2 x 1.8 m l 10x R ea ction Buffer 1.5 ml 50 mm Mg Cl 2 5 x 5 00 µl R ed Taq DN A Po lymerase, 1 U/µ l 5 x 1.8 ml 10x Rea ction Buffer 5 x 1.5 m l 50 mm Mg Cl 2 10 x 5 00 µ l R ed Taq DN A Pol ymera se, 1 U/µl 2 x 1.8 m l 10x R ea ction Buffer 1.5 ml 50 mm Mg Cl 2 10 x 5 00 µ l R ed Taq DN A Pol ymera se, 1 U/µl 10 x 1.8 m l 10x R ea ction Buffer 10 x 1.5 ml 5 0 mm MgC l 2 High-throughput, simplified PCR setup with easy identification of reaction mixtures containing enzyme Routine amplification to 4 kb and TA cloning Red Taq DNA Polymerase Storage Buffer allows mixtures containing the enzyme to be identified High product yields and robustness Exceptionally pure Taq DNA Polymerase for both routine and demanding PCR applications Taq DNA Polymerase, 5 U/µl, in Storage Buffer: 20 mm Tris-HCl (ph 8.0 at 25 C), 30 mm KCl, 0.1 mm DTT, 0.5% Tween 20, 50% (v/v) glycerol, red dye 10x Reaction Buffer: 670 mm Tris-HCl (ph 8.8 at 25 C), 166 mm (NH 4) 2SO4, 4.5% Triton X-100, 2 mg/ml gelatin biotechrabbit Red Taq DNA Polymerase is a first -choice colored, thermostable DNA polymerase f or simplifying PCR setup. This red-colored enzyme formulation performs in all respects like colorless Taq DNA Polymerase formulations and ensures high product yields from various templates. Additionally, reaction mixtures that contain the enzyme are easily identified. Taq DNA polymerase is a thermostable highly processive 5' 3' DNA polymerase that has low 5 ' 3' exonuclease activity and lacks 3 ' 5' exonuclease (proofreading) activity. The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products, allowing easy cloning of PCR products into vectors with T overhangs. UNIT DEFINITION One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dntp into acid - insoluble form in 30 minutes at 72 C in the presence of the reaction buffer. Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay Red PCR Master Mix, 2x BR BR rea ction s 500 r ea cti ons 2 x 1.25 ml R ed PCR Ma ster Mix, 2x 10 x 1.25 m l R ed PCR Master Mix, 2 x High throughput, simplified PCR setup with easy identification of reaction mixtures containing the enzyme and dntps Routine amplification up to 4 kb TA cloning Exceptionally pure Taq DNA Polymerase and highest quality dntps in a 2x Red PCR Master Mix formulation High product yields and robustness in a wide application range Red PCR Master Mix, 2x: Proprietary composition, contains all necessary PCR reagents, including 3 mm MgCl2 (1x has 1.5 mm MgCl2) biotechrabbit Red PCR Mast er Mix is a perfect choice for simplified reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the colored formulation provides the possibility to identify the reactions w hich already contain the enzyme and dntps. This red-colored master mix performs in all respects like colorless PCR Master Mix and ensures high product yields from various templates. It contains highly purified recombinant biotechrabbit Taq DNA polymerase, highest quality dntps and optimal PCR buffer reagents; thus, only template DNA, PCR primers and PCR-grade water need to be added. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Red PCR Master Mix: Functional PCR Assay 10 Bulk and custom formulations available Please contact oem@biotechrabbit.com

11 Hot Start PCR Product Selection Guide Hot Start PCR is a more sensitive technique than standard PCR that allows amplification of low-abundance targets and single-copy genes while reducing PCR background problems. Thermophilic DNA polymerases are unfortunately active at room temperature, which can result in amplification of unspecific targets due to random primer annealing events. Hot -start enzymes are completely inactive during room -temperature reaction setup and become active only after heating. Thus, misprimed amplification products and primer dimer formation do not occur, diminishing PCR background. Hot-start enzymes are typically created with one of the following techniques: chemically, by the addition of thermo-labile blocking groups, or with binding by one or more Taq-specific antibodies. Both of these techniques prevent enzyme activity until the blockage is disrupted at high temperature. Usually hot-start modification does not affect PCR amplification: after thermal reactivation, hot-start Taq DNA polymerases perform similarly to unmodified Taq with respect to processivity, fidelity and deoxyribonucleot idyl transferase activity, allowing TA cloning. Hot-start PCR master mixes containing all necessary PCR reagents are offered for simplified reaction setup. Colored hot-start PCR buffers are available for direct loading of PCR products on to gels. Prod uct Main featur e Main PCR applicati on Sim p lifi ed setup Opti mi zation freed om Direct ge l load ing Colore d enzyme mixt ures Fide lit y Maxi mu m prod uct len gth TA clonin g Hot Star t Taq DNA Polym erase, 5 U / µl An tibo dy -based Ho t Start Ta q DNA Pol ym erase sup pli ed wi th PCR Boos ter for hig her PCR sp ecifi ci ty a nd b etter yi eld High -s en si tiv ity, hotstart Typi cal for Taq 3 5 k b Hot Star t P CR Master Mi x, 2x An tibo dy -based Ho t Start Ta q DNA Pol ym erase wi th a ll PCR rea gents High -throu ghput, hig h -sp ecifi ci ty, ho t- start Typi cal for Taq 3 5 k b Red Hot Start DNA Polym erase, 5 U / µl An tibo dy -based Ho t Start Ta q DNA Polymerase in a red - color ed S torage Buffer for easy visual ization of mix es con tai nin g the en zyme High -s en si tiv ity, ho t- start wi th easy identifi catio n of reactio ns con taini ng en zyme Typi cal for Taq 3 5 k b Red Hot Start PCR Mast er Mix, 2 x An tibo dy -based Ho t Start Ta q DNA Pol ym erase and all PCR rea gents in R ed Buffer for easy visual ization of mix es con tai nin g the en zyme High -throu ghput, high-specificity hot - start wi th easy identifi catio n of reactio ns con taini ng en zym e Typi cal for Taq 3 5 k b Green Hot Start PC R Master Mi x, 2x An tibo dy -based Ho t Start Ta q DNA Pol ym erase and all PCR rea gents in Green R eactio n Buffer for d irect loadin g o nto gels after the PCR High -throu ghput, hig h -sp ecifi ci ty, ho t- start for dir ect loadin g o n gels Typi cal for Taq 3 5 k b info@biotechrabbit.com 11

12 Hot Start Taq DNA Polymerase, 5 U/µl BR BR U 500 U 20 µl H ot S tart Taq DNA Pol ym erase, 5 U /µ l 1.8 ml 10x R eactio n Buffer 1.5 ml 50 mm Mg Cl ml 5x PCR Bo os ter 100 µl Hot S tart Ta q DNA Pol ym erase, 5 U /µ l 2 x 1.8 m l 10x R ea ction Buffer 1.5 ml 50 mm Mg Cl 2 2 x 1.8 m l 5 x PCR Boo ster High-specificity, hot-start amplification up to 4 kb Amplification of low-copy-number targets RT-PCR TA cloning High PCR specificity and sensitivity Hot Start Taq DNA Polymerase for demanding, sensitive PCR applications and high yields Taq DNA Polymerase with bound antibody, in proprietary Storage Buffer 10x Reaction Buffer: 670 mm Tris-HCl (ph 8.8 at 25 C), 166 mm (NH 4) 2SO4, 4.5% Triton X-100, 2 mg/ml gelatin Hot Start PCR Master Mix, 2x BR BR rea ction s 500 r ea cti ons 2 x 1.25 ml Hot S tart PCR Ma ster M ix, 2x 10 x 1.25 m l Ho t Star t PCR Master Mix, 2x High-specificity, high-throughput, hot-start amplification up to 4 kb Amplification of low-copy-number targets TA cloning Highest PCR specificity and sensitivity without prolonged reactivation Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dntps in a 2x Hot Start PCR Master Mix Hot Start PCR Master Mix, 2x: Proprietary composition, contains all necessary hot-start PCR reagents, including 3 mm MgCl2. (1x has 1.5 mm MgCl2) biotechrabbit Hot Start Taq DNA polymerase is a first -choice hot-start PCR enzyme for all demanding PCR applications. The enzyme ensures high product yields with low background; without primer dimer formation and nonspecific priming. The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody whi ch is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation step s. The PCR specificity can be further increased in some cases by addition of the supplied 5x PCR Booster. The optional use of 5x PCR Booster improves PCR results in many cases, including impure template or low template amounts. Taq DNA polymerase is a thermostable highly processive 5' 3' DNA polymerase that has low 5 ' 3' exonuclease activity and lacks 3 ' 5' exonuclease (proofreading) activity. The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products allowing easy cloning of PCR products into vectors with T overhangs. UNIT DEFINITION One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dntp into acid - insoluble form in 30 minutes at 72 C in the presence of the reaction buffer. Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay biotechrabbit Hot Start PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for low -background, highthroughput PCR of kb DNA targets. The 2x Hot Start PCR Master Mix contains pure biotechrabbit Hot Start Taq DNA polymerase, highest quality dntps and optimal PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added. The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation step s. Thus, primer dimers and miss-priming do not occur. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Hot Start PCR Master Mix: Functional PCR Assay 12 Bulk and custom formulations available Please contact oem@biotechrabbit.com

13 Red Hot Start DNA Polymerase, 5 U/µl BR BR BR U 500 U 5 x 5 00 U 20 µl Red Ho t Star t DN A Pol ym erase, 5 U /µ l 1.8 ml 10x R eactio n Buffer 1.5 ml 50 mm Mg Cl ml 5x PCR Bo os ter 100 µl R ed H ot S tart DN A Pol ymera se, 5 U /µ l 2 x 1.8 m l 10x R ea ction Buffer 1.5 ml 50 mm Mg Cl 2 2 x 1.8 m l 5 x PCR Boo ster 5 x 1 00 µl Red Ho t Star t DN A Pol ym er ase, 5 U /µ l 5 x 1.8 ml 10x Rea ction Buffer 5 x 1.5 m l 50 mm Mg Cl 2 5 x 1.8 ml 5x PCR Boo s ter High-throughput, simplified PCR setup with easy identification of reaction mixtures that contain enzyme High specificity, hot-start amplification up to 4 kb Amplification of low-copy-number targets TA cloning Red Hot Start DNA Polymerase Storage Buffer allows mixtures containing the enzyme to be identified Highest PCR specificity and sensitivity without prolonged reactivation Red Enzyme Storage Buffer: Proprietary composition, 5 U/µl Hot Start Taq DNA Polymerase in a red-colored Storage Buffer 10x Reaction Buffer: 670 mm Tris-HCl (ph 8.8 at 25 C), 166 mm (NH 4) 2SO4, 4.5% Triton X-100, 2 mg/ml gelatin biotechrabbit Red Hot Start DNA Polymerase is a first -choice hot-start enzyme ensuring high product yields with low background and simplified PCR setup. This red -colored enzyme formulation performs in all respects like colorless Hot Start Taq DNA Polymerase and ensures low -background, high-specificity amplification without primer dimer formation and miss-priming. Additionally, reaction mixtures containing the enzyme are easily identified due to the red-colored enzyme Storage Buffer. The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps. Thus, primer dimer formation and miss-priming do not occur. The PCR specificity can be further increased in some cases by addition of the supplied 5x PCR Booster. The optional use of 5x PCR Booster improves PCR results in many cases, including impure template or low template amounts. Taq DNA polymerase is a thermostable, highly processive 5' 3' DNA polymerase that has low 5 ' 3' exonuclease activity and lacks 3' 5' exonuclease (proofreading) activity. The enzyme also exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products allowing easy cloning of PCR products into vectors with T overhangs. UNIT DEFINITION One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dntp into acid - insoluble form in 30 minutes at 72 C in the presence of the reaction buffer. Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay info@biotechrabbit.com 13

14 Red Hot Start PCR Master Mix, 2x BR BR rea ction s 500 r ea cti ons 2 x 1.25 ml R ed Ho t S tart PCR Master Mix, 2x 10 x 1.25 m l R ed Ho t S tart PCR Master Mix, 2x High-throughput, simplified PCR setup with easy identification of the reaction mixtures that contain enzyme Hot-start PCR amplification up to 4 kb Amplification of low-copy-number targets TA cloning Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dntps in a red -colored master mix for simplified reaction setup Highest PCR specificity and sensitivity without prolonged reactivation Red Hot Start PCR Master Mix, 2x: Proprietary composition, contains all necessary PCR reagents, including 3 mm MgCl2 (1x has 1.5 mm MgCl2) and the red dye biotechrabbit Red Hot Start PCR Master Mix is a perfect choice for simplified hot-start PCR setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. Additionally the colored formulation allows the reactions that contain the enzyme and dntps to be identified. This red-colored master mix performs in all respects like colorless Hot Start PCR Master Mix and ensures high product yields from various templates with low background. The 2x Red Hot Start PCR Master Mix contains pure biotechrabbit Hot Start Taq DNA polymerase, highest quality dntps and optimal colored PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added. The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated PCR temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation steps. Thus, primer dimer formation and misspriming do not occur. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Red Hot Start Master Mix: Functional PCR Assay Green Hot Start PCR Master Mix, 2x BR BR rea ction s 500 r ea cti ons 2 x 1.25 ml Gr een Ho t S tart PCR Master Mix, 2x 10 x 1.25 m l Green Ho t S tar t PCR Master Mi x, 2x High-specificity, high-throughput hot-start PCR for direct loading onto gels Amplification of low-copy-number targets TA Cloning Green Hot Start PCR Master Mix formulation allows direct loading onto gels after PCR Highest PCR specificity and sensitivity without prolonged reactivation Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dntps in a 2x Hot Start PCR Master Mix Green Hot Start PCR Master Mix,2x: Proprietary composition contains all necessary PCR reagents, including 3 mm MgCl2 (1x has 1.5 mm MgCl2) and green and yellow electrophoresis dyes biotechrabbit Green Hot Start PCR Master Mix is a perfect choice for a fast reaction setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for lowbackground, high-throughput PCR amplification of kb DNA targets. Additionally the special formulation allows reactions to be loaded directly onto gels after amplification without adding additional loading dye. The 2x Hot Start PCR Master Mix contains pure biotechrabbit Hot Start Taq DNA polymerase, highest quality dntps and optimal Green PCR Buffer reagents; thus, only template, PCR primers and PCR-grade water are added. Green Reaction Buffer also allows mixtures containing the enzyme to be identified. The Hot Start Taq Polymerase is inactive during reaction setup due to the bound antibody, which is quickly released at elevated temperatures, ensuring the enzyme is active only during PCR. There is no need for prolonged heating or denaturation step s. Thus, the primer dimer formation and miss-priming do not occur. Green Hot Start PCR Master Mix contains two dyes (blue and yellow) that separate during electrophoresis, allowing migration progress to be monitored. Reactions with Green Hot Start PCR Master Mix have sufficient density for direct loading onto agarose gels. The dyes absorb between nm, making standard A 260 readings to determine DNA concentration unreliable. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Green Hot Start PCR Master Mix: Functional PCR Assay 14 Bulk and custom formulations available Please contact oem@biotechrabbit.com

15 High Fidelity and Long Range PCR Product Selection Guide The PCR accuracy achievable with Taq DNA Polymerase (~one error per 45,000 nucleotides incorporated) is not always sufficient. For demanding cloning, sequencing and expression applications, polymerases w activity) are preferred. These proofreading enzymes can reduce PCR error rates by 10 fold or more, depending on the polymerase. Pfu DNA Polymerase, originally isolated from Pyrococcus furiosus, is one of the most commonly used proofreading enzymes. The unmodified form exhibits approximately 8 10 times higher accuracy than Taq DNA Polymerase. PCR fidelity, however, depends greatly on reaction conditions, including ph, magnesium concentration, bal ance of dntps, base composition of the template and purity of the components. When working with Pfu DNA polymerase, be aware that its might cause primer degradation. Generally, primers for use with the Pfu enzyme should be slightly longer than those for Taq, and the enzyme should always be the last reagent added to the reaction mixture. Pfu is slower than Taq and elongation of 1 kb req uires 2 minutes, whereas Taq polymerase typically requires 1 minute. The synergy of Taq DNA polymerase and a proofreading DNA polymerase is used to generate longer PCR products with higher accuracy than that of Taq alone. Such enzymes blends can be used for the amplification of templates of up to kb in length and of GC-rich templates. In the latter case, different additives ( such as DMSO, formamide or betaine) are helpful to enhance the denaturation of GC -rich templates for higher yields. Prod uct Main featur e Main a pplicati on Fide lit y Maxi mu m prod uct len gth GC-r ich temp lates Sim p lifi ed setup Opti mi zation freed om Blunt Clon in g Use of du TP Hi gh yie ld Pfu DNA Polym erase, 2.5 U/ µl Ten ti m es mor e High -fid eli ty accura te than Taq DNA Pol ym erase; suppli ed wi th PCR Help er for b etter resul ts ~1 0x hig her tha n Taq 3-5 kb No t re c o m m e nded Pfu PC R Master Mi x, 2x Inclu d es Pfu DNA Pol ym erase and hig h -fid eli ty PCR reag en ts High - throu g hpu t, hig h -fid eli ty ~1 0x hig her tha n Taq 3 kb No t re c o m m e nded Lon g and Hi gh Fide lit y P CR Enzym e Mix, 2.5 U/ µl A b lend of ther mos tabl e DNA Pol ym erases for lon g-rang e and hi gh -fid eli ty PCR, wi th PCR Help er for b etter results with GCrich templa tes Lon g-range, GC-ri ch templa tes Similar to Pfu >3 0 kb requir es blun tin g No t re c o m m e nded Lon g Ran ge PCR Mast er Mix, 2x Inclu d es a bl end of thermo sta bl e DNA Pol ym erases and r eag en ts for long -rang e a nd hig h -fid eli ty PCR and GC-rich templa tes High - throu g hpu t, long -rang e, GC-ri ch templa tes Similar to Pfu >3 0 kb requir es blun tin g No t re c o m m e nded info@biotechrabbit.com 15

16 Pfu DNA Polymerase, 2.5 U/µl BR BR U 500 U High-fidelity PCR Blunt cloning of PCR products Site-directed mutagenesis 40 µl Pfu DNA Pol ym erase, 2.5 U /µl 0.5 ml 10x Pfu R eactio n Buff er ml 5x PCR Help er 200 µ l Pfu DN A Pol ymera se, 2.5 U /µ l 2.4 ml 1 0x Pfu R ea ction Buffer 1.2 ml 5 x PCR H elp er Accurate PCR for demanding applications Approximately 10 times higher accuracy than that of Taq DNA Polymerase 2.5 U/µl Pfu DNA Polymerase in Storage Buffer: 50 mm Tris-HCl (ph 8.0 at 25 C), 1 mm DTT, 0.1 mm EDTA, 0.1% (v/v) Nonidet P-40, 1 mm PMSF, 0.1% (v/v) Tween 20, 50% (v/v) glycerol 10x Pfu Reaction Buffer: 500 mm Tris-HCl, 140 mm (NH 4) 2SO4, 17.5 mm MgCl2, ph 9.1 at 25 C. biotechrabbit Pfu DNA Polymerase is a highly purified recombinant thermostable proofreading DNA polymerase. Pfu DNA Polymerase exhibits approximately 10 times higher accuracy than Taq DNA Polymerase. The enzyme catalyzes template -dependent nucleotide polymerization in the 5' 3' direction. Additionally it exhibits 3' 5' exonuclease (proofreading) activity that allows the correction of nucleotide incorporation errors, thereby increasing fidelity and accuracy of PCR. The enzyme has no 5' 3' exonuclease activity and no detectable reverse transcriptase activity. For the most demanding applications, the supplied 5x PCR Helper can be optionally used for improving results when using templates with GC-rich sequences and complex structures. The use of dutp and ditp is not recommended with Pfu, because Pfu binding to DNA templates with uracil and hypoxanthine stalls DNA synthesis. Pfu DNA Polymerase produces blunt -end PCR products for blunt cloning. UNIT DEFINITION One unit catalyzes the incor poration of 10 nmol of dntps into a polynucleotide fraction in 30 minutes at 72 C. Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay Pfu PCR Master Mix, 2x BR BR rea ction s 500 r ea cti ons High-throughput, high-fidelity PCR Blunt cloning of PCR products 2 x 1.25 ml Pfu PCR Master Mi x, 2x ml 5x PCR Help er 10 x 1.25 m l Pfu PC R Master Mi x, 2x 1.2 ml 5 x PCR H elp er Pure Pfu DNA Polymerase and highest quality dntps in a 2x Pfu PCR Master Mix formulation Accurate PCR for demanding applications Approximately 10 times higher accuracy than that of Taq DNA Polymerase Pfu PCR Master Mix, 2x: Proprietary composition contains all necessary PCR reagents biotechrabbit Pfu PCR Master Mix is a perfect choice for fast high-fidelity PCR setup that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for routine high -throughput, highfidelity amplification of targets up to 3-4 kb in size. The 2x Pfu PCR Master Mix contains Pfu DNA polymerase, highest quality dntps and optimal PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added. For the most demanding applications, the supplied 5x PCR Helper can be optionally be used to improve results when using templates with GC -rich sequences and complex structures. Pfu DNA Polymerase exhibits approximately 10 times higher accuracy compared to Taq DNA Polymerase. The use of dutp and ditp is not recommended with Pfu, because Pfu binding to DNA templates with uracil and hypoxanthine stalls DNA synthesis. Pfu DNA Polymerase produces blunt -end PCR products suitable for blunt cloning. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Pfu PCR Master Mix: Functional PCR Assay 16 Bulk and custom formulations available Please contact oem@biotechrabbit.com

17 Long And High Fidelity PCR Enzyme Mix, 2.5 U/µl BR BR U 500 U 40 µl Long and Hi g h Fid el ity PCR Enz yme Mix, 2.5 U/µl 0.5 ml 10x HF R eactio n Buff er ml 5x PCR Help er 200 µ l Lo ng a nd Hig h Fid eli ty PCR En z ym e Mix, 2.5 U/µl 2.4 ml 1 0x H F R ea ction Buffer 1.2 ml 5 x PCR H elp er High-fidelity, long-range PCR Amplification of GC-rich templates Accurate, long-range amplification up to 30 kb Accurate amplification of GC -rich templates 2.5 U/µl thermophilic polymerase blend in storage buffer: 20 mm Tris-HCl (ph 8.0 at 25 C), 100 mm KCl, 1 mm DTT, 0.1 mm EDTA, 0.1% (v/v) Nonidet P -40, 1 mm PMSF, 0.1% (v/v) Tween 20, 50% (v/v) glycerol 10x HF Reaction Buffer: 500 mm Tris-HCl, 140 mm (NH4) 2SO4, 17.5 mm MgCl2, ph 9.1 at 25 C. biotechrabbit Long and High Fidelity PCR Enzyme Mix is a first-choice enzyme blend for amplification of targets up to 40 kb in size with higher accuracy than that of Taq DNA Polymerase. This specially designed blend of thermophilic polymerases is well suited for amplification of targets that are GC-rich and have complex structures. For the most demanding applications, the supplied 5x PCR Helper can be optionally used to improve results when using templates with GC -rich sequences and complex structures. Long and High Fidelity PCR Enzyme Mix produces a mixture of A-tailed and blunt -end PCR products: it is advisable to blunt products before cloning into blunt -end vectors. UNIT DEFINITION One unit catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotid e fraction (adsorbed on DE-81) in 30 minutes at 72 C. Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Functional PCR Assay Long Range PCR Master Mix, 2x BR BR rea ction s 500 r ea cti ons 2 x 1.25 ml Lon g Ra ng e PCR Master Mix, 2x ml 5x PCR Help er 10 x 1.25 m l Lon g Ra ng e PCR Master Mix, 2x 1.2 ml 5 x PCR H elp er High-throughput, long-range PCR Accurate amplification of more than 30 kb Blunt cloning of PCR products Long-range and accurate amplification of more than 30 kb A blend of thermophilic polymerases and highest quality dntps in a convenient 2x master mix formulation Long Range PCR Master Mix, 2x: Proprietary composition contains a blend of thermophilic DNA polymerases and all necessary PCR reagents biotechrabbit Long Range PCR Master Mix is a perfect choice for fast reaction setup for long-range PCR that reduces the time required for calculation and pipetting and eliminates the need for buffer optimization. It is designed for amplification of targets up to 30 kb in size. The master mix works well with GCrich templates and amplifies DNA with a higher fidelity than that of Taq DNA Polymerase. The Long Range PCR Master Mix contains a blend of thermophilic polymerases, highest quality dntps and optimal PCR buffer reagents; thus, only template, PCR primers and PCR-grade water are added. For the most demanding applications, the supplied 5x PCR Helper can be optionally used to improve results when using templates with GC -rich sequences and complex structures. Long Range PCR Master Mix produces a mixture of A-tailed and blunt-end PCR products: it is advisable to blunt products before cloning into the blunt -end vector. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay Long Range PCR Master Mix: Functional PCR Assay info@biotechrabbit.com 17

18 Reverse Transcription and RT-PCR Product Selection Guide The basic difference between PCR and RT-PCR is the template: DNA is the PCR template and RNA is the RT-PCR template. The first step in RT-PCR is the synthesis of single-stranded antisense DNA from an RNA template. This process employs reverse transcriptases (RNA-dependent DNA polymerases). The Moloney-Murine Leukemia Virus (M-MuLV) reverse transcriptase is a classic example of such an enzyme. Mutations have been introduced into reverse transcriptase genes to increase performance. biotechrabbit offers a modified reverse transcriptase, RevertUP Reverse Transcriptase, which is a proprietary modification of the M-MuLV reverse transcriptase with point mutations in polymerase and RNAse H domains. These mutations ensure elevated cdna synthesis efficiency and eliminate RNase H activity, cdna can be synthesized by reverse transcriptase either using random primers, oligo -dt primers or sequence -specific primers. Special care should be taken to prevent RNA templates from degradation by pervasive RNAses. Therefore it is always a safe approach to use RNAse inhibitors in all reactions involving RNA. RT-PCR can be performed in one step or two steps. Two -step RT-PCR includes two different experiments, first the reverse transcription of RNA template is performed and then the PCR amplification of the cdna template is performed in another tube. One - step RT-PCR combines both reactions in one tube : cdna is synthesized from RNA and then amplified to generate cdna with specific primers without additional pipetting steps. Taq DNA Polymerase and its hot-start versions are the main enzymes used for cdna amplification in both the one -step and two-step RT-PCR. Product Application Main feature One Step RT- PCR Kit One-step RT-PCR A blend of efficient thermostable reverse transcriptase and proprietary Ribonuclease Inhibitor provides high cdna yields. Unique Hot Start Taq DNA Polymerase in a mix with high -quality dntps and PCR enhancers allows sensitive, low-background amplification. RevertUP Reverse Transcriptase Reliable synthesis of, two-step RT-PCR A proprietary M -MuLV reverse transcriptase engineered by point mutations in polymerase and RNAse H domains that ensure efficient cdna synthesis with no RNAse H activity, allowing succe MMuLV Reverse Transcriptase cdna synthesis, twostep RT-PCR A classical, exceptionally pure reverse transcriptase supplied with Reaction Buffer. RNAse Inhibitor Prevention of RNA degradation by RNAses A recombinant, non-competitive inhibitor of pancreatic -type ribonucleases such as RNAse A, RNAse B, and RNAse C. 18 Bulk and custom formulations available Please contact oem@biotechrabbit.com

19 One Step RT-PCR Kit BR BR BR rea ction s 100 rea ction s 500 r ea cti ons One-step RT-PCR Cloning Expression-level determination Virus detection µl On e S tep Mi x, 2x 125 µl RT RI Bl end, 2 0x 2 x µl On e S tep Mix, 2x 2 x 12 5 µl R T RI Bl en d, 2 0x 10 x µ l On e Step Mix, 2x 10 x 12 5 µl R T RI Bl en d, 2 0x A blend of efficient reverse transcriptase and proprietary Ribonuclease Inhibitor provides high cdna yields Unique Hot Start Taq DNA Polymerase in a mix with high-quality dntps and PCR enhancers allows sensitive, low-background amplification Successful amplification of GC-rich and complex templates One Step Mix, 2x: Proprietary composition, a mix of Hot Start Taq Polymerase, 6 mm MgCl2, 2 mm dntps, enhancers and stabilizers RT-RI Blend, 20x: Proprietary composition, a blend of efficient reverse transcriptase and unique Ribonuclease Inhibitor biotechrabbit One Step RT-PCR Kit provides an easy and efficient way to perform both reverse transcription of RNA and cdna amplification by PCR in one step. Only RNA template, primers and PCR-grade water are added. The 20x RT RI Blend a blend of an efficient thermostable reverse transcriptase and a proprietary Ribonuclease Inhibitor ensures high yields of cdna. The 2x One Step Mix, which contains unique Hot Start Taq DNA Polymerase, dntps, MgCl 2 and stabilizers in an optimized buffer, provides high PCR product yields with minim al backgrounds even from low-abundance template and difficult targets. The PCR enhancers included in the mix allow efficient amplification of complex templates including GC - or AT-rich sequences. Functional RT-PCR Assay NEW RevertUP Reverse Transcriptase, 100 U/µl BR BR U for 100 rea ction s (20 µl) U for 500 r ea cti ons (20 µl) First-strand cdna synthesis cdna labeling RNA analysis by primer extension 100 µl R ev ertup Reverse Trans crip tase, 100 U /µl 1 ml 5x R ev ertup Buffer 500 µl RevertUP R everse Trans crip tase, 100 U /µl 5 x 1 m l 5x RevertUP Buff er Efficient at high temperatures (up to 55 o C) High-sensitivity cdna synthesis from a few template copies Exceptionally pure enzyme for demanding applications RevertUP Reverse Transcriptase 100 U/µl in Storage Buffer: 50 mm Tris-HCl (ph 7.5 at 25 C), 100 mm NaCl, 0.1 mm EDTA, 10 mm DTT, 0.01% Nonidet P -40, 50% glycerol 5x RevertUP Buffer: Proprietary, contains 25 mm Mg 2+ RevertUP is a proprietary MMuLV reverse transcriptase engineered by point mutations in polymerase and RNase H domains. This ensures efficient cdna synthesis and eliminate RNase H activity, cdna. Reverse transcriptase is a DNA polymerase which utilizes RNA as a substrate and exhibits no measurable proofreading 3 ' 5' exonuclease function. Enzyme performs cdna synthesis by extending a DNA primer annealed to an RNA template or copies ssdna template. UNIT DEFINITION One unit is the amount of enzyme activity that incorporates 1 nmole of dttp into acid insoluble fraction in 10 minutes at 42 C when poly A + RNA and oligo-dt(20) are used as template primer. RNAse Assay Functional Assay Rev ertu P RTas e X R Tase Y P i c ture 1. RT- P C R of th e G3P DH gene ( 500 b p ). c DNA te m p l a te s were s y nth e s i z e d w i th v a ri o us RNAs e H re v e rs e tra ns c ri p ta s e s f ro m G3PD H mr NA ( c o p i e s /re a c ti o n). The re v e rs e tra ns c ri p ti o n re a c ti o n was perf o rm e d with s p e c i f i c re v e rs e pri m e rs a nd 100 U e nz y m e a t 42ºC f o r 20 minute s info@biotechrabbit.com 19

20 MMuLV Reverse Transcriptase, 200 U/µl BR BR U for 50 rea ction s (10 µl) U for 250 rea ction s (10 µl) First-strand cdna synthesis Generation of labeled cdna RNA analysis by primer extension 50 µl MMu LV R ev erse Transcrip tase, 200 U/µl 1 ml 10x MMuLV RT Buffer 250 µl MMu LV R ev erse Tra ns crip tase, 200 U/µl 1 ml 10x MMuLV RT Buffer High yields of first-strand cdna Exceptionally pure reverse transcriptase for demanding applications MMuLV Reverse Transcriptase, 200 U/µl in Storage Buffer: 50 mm Tris-HCl, 150 mm NaCl, 0.1 mm EDTA,1 mm DTT, 0.1% NP-40 Alternative, 50% glycerol, ph 7.6 at 25 C 10x MMuLV RT Buffer: 500 mm Tris-HCl, 750 mm KCl, 30 mm MgCl 2, 100 mm DTT, ph 8.3 at 25 C biotechrabbit MMuLV Reverse Transcriptase is an exceptionally pure DNA polymerase which utilizes RNA as a substrate and exhibits no measurable proofreading 3 ' 5' exonuclease function. This enzyme can perform cdna synthesis by extending a DNA primer annealed to an RNA template, or can copy a single-stranded DNA template. The enzyme is purified from a recombinant E. coli strain carrying the MMuLV reverse transcriptase gene. UNIT DEFINITION One unit is defined as the amount of enzyme required to incorporate 1 nmol of dttp into acid insol uble material in 10 minutes at 37 C using poly ra/oligo -dt as a substrate. Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rdna Contamination Test Non-Specific RNAse Assay RNAse Inhibitor, 40 U/µl BR U 62.5 µ l RN As e In hibi tor, 4 0 U /µl BR U 250 µl RN As e I nhibi tor, 4 0 U/µl Prevention of RNA degradation by Ribonucleases A, B or C during: In vitro transcription and translation cdna synthesis RNA purification and storage Exceptionally pure proprietary Ribonuclease Inhibitor for demanding RNA applications Active under variety of reaction conditions for RNA RNAse Inhibitor, 40 U/µl in Storage Buffer: 20 mm Hepes-KOH, 50 mm KCl, 8 mm DTT, 50% glycerol, ph 7.5 at 25 C biotechrabbit RNAse Inhibitor is an acidic, 52 kda protein that is a potent non-competitive inhibitor of pancreatic -type ribonucleases such as RNase A, RNase B, and RNase C. The enzyme is provided as a fusion of the porcine RNAse Inhibitor gene with a proprietary, 22.5 kda protein tag. The protein is purified from a recombinant E. coli strain carrying the porcine RNAse Inhibitor gene. UNIT DEFINITION One unit is defined as the amount of enzyme required for 50% inhibition of -cyclic monophosphate by 5 ng of RNAse A. Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rdna Contamination Test Non-Specific RNAse Assay 20 Bulk and custom formulations available Please contact oem@biotechrabbit.com

21 Quantitative Real-Time PCR Product Selection Guide The difference between quantitative real-time PCR (qpcr) and end-point PCR is the step at which the PCR product amount is measured. In traditional PCR, product quantitation is performed once at the end of the reaction. In qpcr, the amount of PCR product is measured after every amplification round. qpcr instruments are designed for both PCR c ycling and the detection of signal from fluorescent dyes during amplification. By monitoring the progress of amplification in real time, the point in time when the amplification of the target is detected above background can be identified. The fewer target s present in the reaction, the later the first significant signal is detected. This technique, therefore, is of great advantage when gene -expression differences are analyzed. There are two ways to detect PCR signal with the help of fluorescent dyes: fluorescent dyes that nonspecifically bind to double - stranded DNA or fluorescently labeled, sequence-specific probes. The use of universal fluorescent dyes enables analysis of many targets without having to synthesize sequence -specific probes. However, this method has the disadvantage of detecting all specific and unspecific amplification events as well as primer dimers because the dye binds to all double-stranded DNA. To achieving high PCR specificity, avoiding background and primer dimer formation is critical. The fluorescent -probe method is more precise, as the use of sequence -specific fluorescently labeled probes ensures the detection of the target product only. A variety of probe fluorescence chemistry is currently available. Multiple fluorescent probe s can be used in multiplex qpcr, although the use of specific probes is more expensive than using DNA-binding dyes. QPCR OneStep Kits allow reverse transcription and real-time PCR to be performed in one step. In this case, RNA is the template for qpcr. Some one-step kits for qpcr provide an intercalating dye for probe detection. One -step qpcr kits include separate qpcr and reverse transcription master mixes. Many real-time PCR instruments available on the market require a passive internal reference dye (either ROX or fluorescein) for normalization of the fluorescent signal. The use of passive reference dyes does not interfere with the qpcr. The table below will help you select a qpcr master mix for use on your instrument: Real-time PCR master mixes (for fast or standard mode and for multiplex PCR) Intercalating -dye-based mixes (S YBR Green I, EvaGreen or other), such as QPCR Green Master Mix Nonspecific, detects all d oublestranded DNA Cheaper and faster Probe-based mixes (TaqMan, Molecular Beacon or other), such as QPCR Probe Master Mix Can be used specifically Can be universally More expensive Takes more time to design and synthesize specific probes Passive reference dye ROX Instruments Applied Biosystems: ABI PRISM 7000, 7300, 7900 HT, and StepOne 7500, ROX solution should be provided separately Fluorescein Bio-Rad icycler iq and iq 5 None ROX Fluorescein None Roche LightCycler 480, Roche LightCycler 1.5, 2.0, Bio-Rad SmartCycler, QIAGEN Rotor-Gene 3000, Rotor-Gene 6000, Eppendorf Mastercycler ep REALPLEX and Techne Quantica Applied Biosystems: ABI PRISM 7000, 7300, 7900 HT, and 7500, ROX solution should be provided separately Bio-Rad icycler iq 5, QIAGEN Rotor-Gene 6000, Bio- Rad CFX96 or Roche LightCycler 480. Roche LightCycler 480, Roche LightCycler 1.5, 2.0, Bio-Rad SmartCycler, QIAGEN Rotor-Gene 3000, Rotor-Gene 6000, Eppendorf Mastercycler ep REALPLEX and Techne Quantica for ; for info@biotechrabbit.com 21

22 biotechrabbit qpcr master mixes that do not contain the reference dye in the mix (reference dye can be supplied separately ): Product QPCR Green Master Mix, 2x QPCR Probe Master Mix, 2x Reference dye supplied Application Instruments Roche LightCycler 480, Roche LightCycler 1.5, 2.0, Bio-Rad None ROX, separate Fluorescein, separate None ROX, separate Fluorescein, separate Intercalating-dye-based qpcr Intercalating-dye-based qpcr on ROX instruments Intercalating-dye-based qpcr on fluorescein instruments Probe-based qpcr Probe-based qpcr on ROX instruments Probe-based qpcr on fluorescein instruments SmartCycler, QIAGEN Rotor-Gene 3000, Rotor-Gene 6000, Eppendorf Mastercycler ep REALPLEX and Techne Quantica Applied Biosystems: ABI PRISM 7000, 7300, 7900 HT and ; for 7500, ROX solution should be provided separately Bio-Rad icycler iq 5 Roche LightCycler 480, Roche LightCycler 1.5, 2.0, Bio-Rad SmartCycler, QIAGEN Rotor-Gene 3000, Rotor-Gene 6000, Eppendorf Mastercycler ep REALPLEX and Techne Quantica Applied Biosystems: ABI PRISM 7000, 7300, 7900 HT and StepOne for 7500, ROX solution should be provided separately Bio-Rad icycler iq 5, QIAGEN Rotor-Gene 6000, Bio- Rad CFX96 and Roche LightCycler 480 biotechrabbit qpcr master mixes that contain ROX (low or high concentration) in the master mix: Product Reference dye supplied Company Instrument QPCR SyGreen Master Mix, LRox and QPCR Probe-R Master Mix, LRox QPCR SyGreen Master Mix, HRox and QPCR Probe-R Master Mix, HRox Low-concentration ROX, in mix High concentration ROX, in mix Analytica Jena Applied Biosystems Bio-Rad Cepheid Eppendorf Illumina QIAGEN/Corbett Roche Applied Science Stratagene (Agilent) Takara Techne Analytica Jena Applied Biosystems Cepheid Eppendorf Illumina QIAGEN/Corbett Roche Applied Science Takara Techne qtower 7500, 7500 FAST, Viia 7 icycler, MyiQ, iq 5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 Smartcycler Mastercycler ep REALPLEX, Mastercycler REALPLEX 2S Eco Rotor-Gene 3000, 6000, Q Lightcycler 480, Lightcycler Nano Mx4000P, Mx3000P, Mx3005P Cycler Dice Quantica qtower 7000, 7300,7700,7900, 7900HT, 7900HT FAST, StepOne, StepOne Plus Smartcycler Mastercycler ep REALPLEX, Mastercycler REALPLEX 2S Eco Rotor-Gene 3000, 6000, Q Lightcycler 480, Lightcycler Nano Cycler Dice Quantica biotechrabbit QRT-PCR OneStep Kits: QRT-PCR SyGreen or Probe-R One Step Kits, LROX QRT-PCR SyGreen or Probe-R One Step Kits, HROX Company Instrument Yes Yes Analytica Jena qtower Yes NO Applied Biosystems 7500, 7500 FAST, Viia 7 NO Yes Applied Biosystems Yes NO Bio-Rad Yes Yes Cepheid Smartcycler 7000, 7300,7700,7900, 7900HT, 7900HT FAST, StepOne, StepOne Plus icycler, MyiQ, iq 5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 Yes Yes Eppendorf Mastercycler ep REALPLEX, Mastercycler REALPLEX 2S Yes Yes Illumina Eco Yes Yes QIAGEN/Corbett Rotor-Gene 3000, 6000, Q Yes Yes Roche Applied Science Lightcycler 480, Lightcycler Nano Yes NO Stratagene (Agilent) Mx4000P, Mx3000P, Mx3005P Yes Yes Takara Cycler Dice Yes Yes Techne Quantica 22 Bulk and custom formulations available Please contact oem@biotechrabbit.com

23 QPCR Green Master Mix, 2x Intercalating-dye-based qpcr mixes without reference dye PRODUCT SIZE BR BR BR BR BR BR QPCR Green Master Mi x, 2x QPCR Green Master Mi x, 2x QPCR Green Master Mi x, 2x QPCR Green Master Mi x, 2x w ROX QPCR Green Master Mi x, 2x w ROX QPCR Green Master Mi x, 2x w Fluor es cein 50 reactio ns 200 reactio ns 2000 reactio ns 50 µl) 50 reactio ns 200 reactio ns 200 reactio ns ml QPCR Gr een Master Mi x, 2x 4 x 1.25 m l QPCR Gr een Master Mi x, 2x 50 ml QPCR Green Master Mix, 2x ml QPCR Gr een Master Mi x, 2x ml RO X, 5 µm 4 x 1.25 m l QPCR Gr een Master Mi x, 2x 1 ml R O X, 5 µm 4 x 1.25 m l QPCR Gr een Master Mi x, 2x 0.4 ml Fl uorescei n, 5 00 nm Always ca librat e yo ur instru ment in a separate r un. Do not inc lu de passive refe renc e d yes in the QP CR Maste r Mix. qpcr based on intercalating green dye Exceptionally pure Hot Start Taq DNA Polymerase, a proprietary green dye, and highest quality dntps in a 2x QPCR Green Master Mix formulation Excellent PCR efficiency, correlation coefficient and slope For maximum flexibility, ROX or fluorescein supplied separately QPCR Green Master Mix, 2x: Proprietary composition, contains all necessary qpcr reagents, including proprietary green dye biotechrabbit QPCR Green Master Mix is a fi rst-choice mix for fast and easy real-time PCR setup. It contains all reagents required for real-time PCR and is designed to achieve excellent results in reaction efficiency, correlation coefficients and amplification slope. The master mix contains our pure Hot Start Taq DNA Polymerase, highest quality dntps and buffer reagents, all at optimal concentrations. Only template, primers and PCR-grade water are added. The green intercalating dye allows the universal detection of all PCR products without the use of sequence -specific probes. The dye binds to double -stranded DNA and emits a fluorescent signal upon binding. In qpcr, DNA accumulates and fluorescent signal increases proportionally to the DNA concentration. The excitation and emission m axima of the green dye are compatible with use on any real -time cycler. The master mix has no premixed reference dye therefore can be successfully used with most real-time PCR instruments. For more flexibility, master mix packages with either ROX or fluorescein as passive reference dyes are provided. For green-dye-based amplicon detection, it is important to run a dissociation curve following the real -time PCR. This is necessary because the green dye will detect any double - stranded DNA, including primer dimers, contaminating DNA, and PCR product s from miss-annealed primers. The melting temperature of the amplicon correlates with the maximum of the melting curve profile. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay QPCR Green Master Mix: Functional QPCR Assay Picture 1. High qpcr specificity achieved using QPCR Green Master Mix. The melting curve (GAPDH endogenous control) shows that the maximum (melting temperature of the amplicon) occurs at 81.5 C, there are no contaminating products, which would appear as additional peaks. info@biotechrabbit.com 23

24 QPCR SyGreen Master Mix LRox and HRox, 2x Intercalating-dye-based qpcr mixes with ROX PRODUCT SIZE BR BR BR BR BR BR QPCR S ygreen Master Mi x, LR ox, 2x QPCR S ygreen Master Mi x, LR ox, 2x QPCR S ygreen Master Mi x, LR ox, 2x QPCR S ygreen Master Mi x, HRox, 2x QPCR S ygreen Master Mi x, HRox, 2x QPCR S ygreen Master Mi x, HRox, 2x 100 reactio ns (20 µl) 500 reactio ns (20 µl) 2000 reactio ns (20 µl) 100 reactio ns (20 µl) 500 reactio ns (20 µl) 2000 reactio ns (20 µl) 1 ml QPCR SyGr een Master Mi x, LR ox, 2x 5 x 1 m l QPCR SyGreen Master Mi x, LR ox, 2x 20 x 1 ml Q PCR S ygr een Master Mi x, LR ox, 2x 1 ml QPCR SyGr een Master Mi x, HRox, 2 x 5 x 1 m l QPCR SyGreen Master Mi x, HRox, 2 x 20 x 1 ml Q PCR S ygr een Master Mi x, HRox, 2 x qpcr based on intercalating-green-dye detection instruments requiring ROX as reference dye No PCR inhibition from intercalating dye Rapid extension rate for early CT values Highest sensitivity for increased limit of detection Compatible on all real -time PCR platforms for standard and fast cycling conditions 2x QPCR SyGreen Master Mix LRox and HRox: Proprietary composition, contains all necessary qpcr reagents including proprietary green dye and ROX at a low or high concentration biotechrabbit QPCR SyGreen Master Mixes with ROX in high or low concentration are first-choice products for fast and easy real-time PCR setup. They contain all reagents required for real-time PCR and are designed to achieve excellent results in reaction efficiency, correlation coefficient and slope. QPCR SyGreen Master Mixes use a proprietary intercalating dye which does not inhibit PCR, unlike other popular fluorescent dyes. QPCR SyGreen Master Mixes ca n be used to quantify any DNA template including genomic, cdna and viral sequences. Extremely low-copy-number targets can be detected specifically with high efficiency. Proprietary technology prevents formation of primer dimers and non-specific products leading to improved reaction sensitivity and specificity. Many modern real-time PCR instruments have a fast cycling mode. The same developments that improve the sensitivity and consistency of QPCR SyGreen Master Mixes in standard cycling conditions also all ow industry-leading performance in fast and ultra fast cycling conditions. For more flexibility, ROX is provided either at a high (HRox) or low concentration (LRox). Components tested for absence of ribonuclease, endonuclease and exonuclease activity. Mix tested functionally in qpcr. Choose LRox or HRox Master Mix according to the instrument: Company Instrument LRox HRox Analytica Jena qtower Yes Yes Applied Biosystems Applied Biosystems Bio-Rad 7500, 7500 FAST, Viia 7 Yes NO 7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne, StepOne Plus icycler, MyiQ, iq 5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 NO Yes Yes Cepheid Smartcycler Yes Yes Eppendorf Mastercycler ep REALPLEX, Mastercycler REALPLEX 2S Yes NO Yes Illumina Eco Yes Yes QIAGEN/Corbett Rotor-Gene 3000, 6000, Q Yes Yes Roche Applied Science Stratagene (Agilent) Lightcycler 480, Lightcycler Nano Yes Yes Mx4000P, Mx3000P, Mx3005P Yes NO Takara Cycler Dice Yes Yes Techne Quantica Yes Yes 24 Bulk and custom formulations available Please contact oem@biotechrabbit.com

25 QPCR Probe Master Mix, 2x Probe-based qpcr mixes without reference dye PRODUCT SIZE BR BR BR BR BR BR QPCR Prob e Master Mi x, 2x QPCR Prob e Master Mi x, 2x QPCR Prob e Master Mi x, 2x QPCR Prob e Master Mi x, 2x w R OX QPCR Prob e Master Mi x, 2x w R OX QPCR Prob e Master Mi x, 2x w Flu orescei n 50 rea ction s 200 r eactio ns 2000 rea ction s 50 rea ction s 200 r eactio ns 200 r eactio ns ml QPCR Pro be Master Mi x, 2x 4 x 1.25 m l QPCR Prob e Ma ster M ix, 2x 50 ml QPCR prob e Master Mi x, 2x 1 x ml QPCR Prob e Ma ster M ix, 2x ml RO X, 5 µm 4 x 1.25 m l QPCR Prob e Ma ster M ix, 2x 1 ml R O X, 5 µm 4 x 1.25 m l QPCR Prob e Ma ster M ix, 2x 0.4 ml Fl uorescei n, 500 nm Always ca librat e yo ur instru ment in a separate r un. Do not inc lu de passive refe renc e d yes in the QP CR Maste r Mix. qpcr based on specific probes Exceptionally pure Hot Start Taq DNA Polymerase and highest quality dntps in a 2x QPCR Probe Master Mix formulation Excellent PCR efficiency, correlation coefficient and slope For maximum flexibility, ROX or fluorescein supplied separately QPCR Probe Master Mix, 2x: Proprietary composition, contains all necessary qpcr reagents biotechrabbit QPCR Probe Master Mix is a first -choice mix for fast and easy real-time PCR setup. It contains all reagents required for real-time PCR and is designed to achieve excellent results in reaction efficiency, correlation coefficient and slope. The master mix contains pure Hot Start Taq DNA Polymerase, highest quality dntps and buffer reagents, all at optimal concentrations. Only template, primers and PCR -grade water are added. The master mix has no premixed ref erence dye therefore can be successfully used with most real -time PCR instruments. For more flexibility, master mix packages with either ROX or fluorescein as passive reference dyes are provided. Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay QPCR Probe Master Mix: Functional QPCR Assay Pict ure 1. Unif orm and rep rod uc ible resu lts a chieve d usin g 2 x QP CR Probe Mast er Mi x. Effici en cy of the ampl ifica tio n of huma n GA PDH u sin g l ogarithmi c serial dilu tion s fro m 1 ng to 1 p g on the R G (Corb ett R es ear ch). info@biotechrabbit.com 25

26 QPCR Probe-R Master Mix LRox and HRox, 2x Probe-based qpcr mixes with ROX PRODUCT SIZE BR BR BR BR BR BR QPCR Prob e -R Master Mi x, LRox, 2 x QPCR Prob e -R Master Mi x, LRox, 2 x QPCR Prob e -R Master Mi x, LRox, 2 x QPCR Prob e -R Master Mi x, HRox, 2x QPCR Prob e -R Master Mi x, HRox, 2x QPCR Prob e -R Master Mi x, HRox, 2x 100 rea ction s (20 µl) 500 r ea cti ons (20 µl) 2000 rea ction s (20 µl) 100 rea ction s (20 µl) 500 r ea cti ons (20 µl) 2000 rea ction s (20 µl) 1 ml QPCR Pro be -R Master Mix, LR ox, 2x 5 x 1 m l QPCR Prob e -R Master Mi x, LR ox, 2x 20 x 1 ml Q PCR Prob e -R Master Mi x, LR ox, 2x 1 ml QPCR Pro be -R Master Mix, HRox, 2 x 5 x 1 m l QPCR Prob e -R Master Mi x, HRox, 2 x 20 x 1 ml Q PCR Prob e -R Master Mi x, HRox, 2 x qpcr based on specific probe detection on instruments requiring ROX as reference dye For use with a wide range of probe technologies, including TaqMan, Molecular Beacon and Scorpions probes High efficiency in multiplex reactions Rapid extension rate for early C T values Highest sensitivity for increased limit of detection Compatible on all real -time PCR platforms for standard and fast cycling conditions 2x QPCR Probe-R Master Mix LRox and HRox Proprietary composition, contains all necessary qpcr reagents including ROX at low or high concentration Components tested for absence of ribonuclease, endonuclease and exonuclease activity. Mix tested functionally in qpcr. biotechrabbit QPCR Probe-R Master Mixes with ROX at low or high concentration are first-choice products for fast and easy real-time PCR setup. They contain all reagents required for real-time PCR and are designed to achieve excellent results in reaction e fficiency, correlation coefficient and slope. QPCR Probe-R Master Mixes use proprietary technology that prevents formation of primer dimers to improve reaction sensitivity and specificity. High-throughput screening was used to identify a buffer system that allows efficient amplification from GC -rich and ATrich templates under both fast and standard cycling conditions. For more flexibility, ROX is provided either at a high (HRox) or low concentration (LRox). Choose LRox or HRox Master Mix according to the instrument: Company Instrument LRox HRox Analytica Jena qtower Yes Yes Applied Biosystems Applied Biosystems Bio-Rad 7500, 7500 FAST, Viia 7 Yes NO 7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne, StepOne Plus icycler, MyiQ, iq 5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 NO Yes Yes Cepheid Smartcycler Yes Yes Eppendorf Mastercycler ep REALPLEX, Mastercycler REALPLEX 2S Yes NO Yes Illumina Eco Yes Yes QIAGEN/Corbett Rotor-Gene 3000, 6000, Q Yes Yes Roche Applied Science Stratagene (Agilent) Lightcycler 480, Lightcycler Nano Yes Yes Mx4000P, Mx3000P, Mx3005P Yes NO Takara Cycler Dice Yes Yes Techne Quantica Yes Yes 26 Bulk and custom formulations available Please contact oem@biotechrabbit.com

27 NEW QRT-PCR SyGreen One Step Kits, LRox and HRox PRODUCT SIZE BR BR BR BR BR BR QRT-PCR SyGr een On e Step Kit, HRox, 2x QRT -PCR SyGr een On e Step Kit, HRox, 2x QRT -PCR SyGr een On e Step Kit, HRox, 2x QRT -PCR SyGr een On e Step Kit, LRox, 2 x QRT -PCR SyGr een On e Step Kit, LRox, 2 x QRT -PCR SyGr een On e Step Kit, LRox, 2 x 100 rea ction s (20 µl) 300 r ea ction s (20 µl) 1200 rea ction s (20 µl) 100 rea ction s (20 µl) 300 r ea ction s (20 µl) 1200 rea ction s (20 µl) 1 ml QPCR SyGr een M ix, HRox, 2x 200 µ l R Tase wi th RNA se Inhi bitor, 20x Mi x 3 x 1 ml QPCR SyGr een Mix, HRox, 2 x 3 x 2 00 µ l R Tase wi th RNAs e I n hibi tor, 2 0x Mix 12 x 1 ml QPCR S ygr een Mix, HRox, 2 x 12 x 2 00 µl R Tase with RNAs e I n hibi tor, 2 0x Mix 1 ml QPCR SyGr een M ix, LRox, 2 x 200 µ l R Tase wi th RNA se Inhi bitor, 20x Mi x 3 x 1 ml QPCR SyGr een Mix, LR ox, 2x 3 x 2 00 µ l R Tase wi th RNAs e I n hibi tor, 2 0x Mix 12 x 1 ml QPCR S ygr een Mix, LR ox, 2x 12 x 2 00 µl R Tase with RNAs e I n hibi tor, 2 0x Mix qpcr from RNA templates based on intercalating green dye Highest sensitivity for increased limit of detection Compatible with standard and fast qpcr platforms Thermostable (55 C), extremely active reverse transcriptase and advanced RNAse inhibitor No PCR inhibition from intercalating dye Rapid extension rate for early C T values QPCR SyGreen Mix, HRox or LRox, 2x Proprietary composition, contains all necessary qpcr reagents RTase with RNase Inhibitor, 20x Mix Proprietary reverse transcriptase in a mix with efficient Ribonuclease Inhibitor biotechrabbit QRT-PCR SyGreen One Step Kits with ROX in low or high concentration are first -choice products for fast and easy real-time PCR setup. They contain all reagents required for one-step QRT-PCR and are designed to achieve excellent results in reaction efficiency, correlation coefficient and slope. QRT-PCR SyGreen One Step Kits allow for efficient cdna synthesis and qpcr in a single tube. They can be used to quantify mrna, total RNA and viral sequences. Extremely low-copy-number targets can be detected with high efficiency. Proprietary technology pr events formation of primer dimers and non-specific products leading to improved reaction sensitivity and specificity. QRT-PCR SyGreen One Step Kits can be used not only in standard cycling conditions, they also show excellent performance in fast and ultra -fast cycling conditions. For more flexibility, ROX is provided either at a high (HRox) or low concentration (LRox). Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay QPCR SyGreen Mix: Functional qpcr Assay Choose the LRox or HRox version, depending on the instrument: Company Instrument LRox HRox Analytica Jena qtower Yes Yes Applied Biosystems 7500, 7500 FAST, Viia 7 Yes NO Applied Biosystems Bio-Rad 7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne, StepOne Plus icycler, MyiQ, iq 5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 Cepheid Smartcycler Yes Yes Eppendorf Mastercycler ep REALPLEX, Mastercycler REALPLEX 2S Yes Yes Illumina Eco Yes Yes QIAGEN/Corbett Rotor-Gene 3000, 6000, Q Yes Yes Roche Applied Science Lightcycler 480, Lightcycler Nano Yes Yes Stratagene (Agilent) Mx4000P, Mx3000P, Mx3005P Yes NO Takara Cycler Dice Yes Yes Techne Quantica Yes Yes NO Yes Yes NO info@biotechrabbit.com 27

28 NEW QRT-PCR Probe-R One Step Kits, LRox and HRox PRODUCT SIZE BR BR BR BR BR BR QRT -PCR Probe- R O ne S tep K it, LRox, 2 x QRT -PCR Probe- R O ne S tep K it, LRox, 2 x QRT -PCR Probe- R O ne S tep K it, LRox, 2 x QRT -PCR Probe- R O ne S tep K it, HRox, 2x QRT -PCR Probe- R O ne S tep K it, HRox, 2x QRT -PCR Probe- R O ne S tep K it, HRox, 2x 100 reactio ns (20 µl) 300 reactio ns (20 µl) 1200 reactio ns (20 µl) 100 reactio ns (20 µl) 300 reactio ns (20 µl) 1200 reactio ns (20 µl) 1 ml QPCR Pro be -R Mix, LRox, 2 x 200 µ l R Tase wi th RNA se Inhi bitor, 20x Mi x 3 x 1 ml QPCR Pro be -R Mix, LR ox, 2x 3 x 2 00 µ l R Tase wi th RNAs e I n hibi tor, 2 0x Mix 12 x 1 ml QPCR Prob e -R Mix, LR ox, 2x 12 x 2 00 µl R Tase with RNAs e I n hibi tor, 2 0x Mix 1 ml QPCR Pro be -R Mix, HRox, 2x 200 µ l R Tase wi th RNA se Inhi bitor, 20x Mi x 3 x 1 ml QPCR Pro be -R Mix, LR ox, 2x 3 x 2 00 µ l R Tase wi th RNAs e I n hibi tor, 2 0x Mix 12 x 1 ml QPCR Prob e -R Mix, LR ox, 2x 12 x 2 00 µl R Tase with RNAs e I n hibi tor, 2 0x Mix qpcr from RNA templates based on specific probes Highest sensitivity for increased limit of detection Compatible with standard and fast qpcr platforms Thermostable (55 C), extremely active reverse transcriptase and advanced RNAse inhibitor Rapid extension rate for early C T values QPCR Probe-R Mix, HRox or LRox, 2x Proprietary composition, contains all necessary qpcr reagents RTase with RNAse Inhibitor, 20x Mix Proprietary reverse transcriptase in a mix with efficient Ribonuclease Inhibitor biotechrabbit QRT-PCR Probe-R One Step Kits with ROX in low or high concentration are first -choice products for fast and easy real-time PCR setup. They contain all reagents required for one -step QRT-PCR and are designed to achieve excellent results in reaction efficiency, correlation coefficient and slope. QRT-PCR Probe-R One Step Kits allow for efficient cdna synthesis and qpcr in a single tube. They can be used to quantify mrna, total RNA and viral sequences. Extremely low-copy-number targets can be detected with high efficiency. Proprietary technology prevents formation of primer dimers and non-specific products leading to improved reaction sensitivity and specificity. QRT-PCR Probe-R One Step Kits can be used not only in standard cycling conditions, they also show excellent performance in fast and ultra -fast cycling conditions. For more flexibility, ROX is provided either at a high (HRox) or low concentration (LRox). Components: Endodeoxyribonuclease Assay Exodeoxyribonuclease Assay Ribonuclease Assay QPCR Probe-R Mix: Functional qpcr Assay Choose the LRox or HRox version, depending on the instrument: Company Instrument LRox HRox Analytica Jena qtower Yes Yes Applied Biosystems 7500, 7500 FAST, Viia 7 Yes NO Applied Biosystems Bio-Rad 7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne, StepOne Plus icycler, MyiQ, iq 5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 Cepheid Smartcycler Yes Yes Eppendorf Mastercycler ep REALPLEX, Mastercycler REALPLEX 2S Yes Yes Illumina Eco Yes Yes QIAGEN/Corbett Rotor-Gene 3000, 6000, Q Yes Yes Roche Applied Science Lightcycler 480, Lightcycler Nano Yes Yes Stratagene (Agilent) Mx4000P, Mx3000P, Mx3005P Yes NO Takara Cycler Dice Yes Yes Techne Quantica Yes Yes NO Yes Yes NO 28 Bulk and custom formulations available Please contact oem@biotechrabbit.com

29 Direct PCR / Crude Sample PCR Direct PCR from crude samples without DNA template purification is an advantageous, time- and cost-saving technique for fast detection of targets of interest. It means amplification directly from blood, soil, food samples and tissues without performing template purification procedure. Direct PCR employs robust, inhibitor-resistant thermophilic polymerases that enable amplification from crude samples containing inhibitors. However, direct PCR might not always be successful or result in high yields and it frequently requires optimization. Therefore, it is important to follow the suggested protocols carefully and not to overload the reaction with template sample to minimize the inhibitory effects of impurities. Biotechrabbit prov ides the robust BlooDirect DNA Polymerase which is used for direct PCR from blood samples or tissue homogenates without the need to purify template DNA. BlooDirect DNA Polymerase, 4.2 U/µl BR BR U 1000 U 100 µl Bloo Direct DN A Pol ymera se, 4.2 U /µl 1 ml 5x Bloo Direct Rea ction Buffer 240 µl Bloo Dir ect DN A Po lymerase, 4.2 U /µl 2x 1ml 5 x Bloo Dir ect R eactio n Buff er Direct PCR from blood [2] or other crude samples Inhibition-resistant Hot Start DNA Polymerase for direct PCR from blood or soil samples without template purification Excellent performance in a wide application range : excellent results with up to 40% whole blood as well as inhibitors in soil and foods and intercalating dyes BlooDirect DNA Polymerase, 4.2 U/µl in Storage Buffer: 10 mm Tris-HCl, 100 mm KCl, 1 mm DTT, 0.1 mm EDTA, 0.3% Brij-58, 50% glycerol, ph 7.4 at 25 C 10x BlooDirect Reaction Buffer: 250 mm Tris-HCl, 80 mm (NH4) 2SO4, 7.5 mm MgCl2, 0.125% Brij 58, ph 9.2 at 25 C biotechrabbit BlooDirect DNA Polymerase is an engineered DNA polymerase providing excellent results without the need for template purification. The enzyme retains robust PCR function in the presence of 40% whole blood, is resistant to the inhibitors present in such complex samples as serum, soil, inhibitory foods, and fluorescent dyes. A series of mutations provides an effective hot-start function which ensures minimized background and eliminates primer dimer formation. The enzyme lacks both 5' 3' and 3' 5' exonuclease activities [1, 2]. The enzyme exhibits deoxynucleotidyl transferase activity that results in the addition of an extra A overhang at the 3' ends of PCR products allowing easy cloning of PCR products into vectors with T overhangs. The protein is purified from E. coli strain carrying a modified recombinant Taq DNA polymerase gene from the strain Thermus aquaticus YT-1. REFERENCES 1. Ker me kchi ev, M.B., et a l. ( 20 03) Nu cle ic Aci ds Res., 31, Ker me kchi ev, M.B., et a l. ( 20 09) Nu cle ic Aci ds Res., 37(5):e 40 E pub. UNIT DEFINITION One unit is defined as the amount of enzyme that will incorporate 10 nmol of dntp into acid -insoluble material in 30 minutes at 75 C. Single-Stranded Exonuclease Assay Double-Stranded Exonuclease Assay Double-Stranded Endonuclease Assay E. coli 16S rdna Contamination Test Picture 1. Performance of BlooDirect DNA Polymerase. Direct PCR of hepatitis B virus from whole blood samples in five serial dilutions. BlooDirect DNA Polymerase detected the test target with similar efficiency from purified DNA and whole blood. Typically, commonly available Taq DNA Polymerases produce no products in direct PCR, and are highly dependent on DNA purification prior to PCR. info@biotechrabbit.com 29

30 dntp Mixes and Sets dntp Mixes and dntp/dutp Mixes PRODUCT SIZE BR mm dn TP Mi x ( 10 mm each) 0.2 m l BR mm dn TP Mi x ( 10 mm each) 1 ml BR mm dn TP Mi x ( 10 mm each) 5 x 0.2 ml BR mm dn TP Mi x ( 10 mm each) 5 x 1 m l BR mm dn TP Mi x ( 10 mm each) 25 x 1 ml BR mm dn TP Mi x (2 mm each) 1 ml BR mm dn TP Mi x (2 mm each) 5 x 1 m l BR mm dn TP Mix (2.5 mm ea ch) 1 ml BR mm dn TP Mix (2.5 mm ea ch) 5 x 1 m l BR mm dn TP M ix ( 25 mm ea ch) 0.5 ml BR mm dn TP M ix ( 25 mm ea ch) 1 ml BR dntp/du TP Mi x (2 mm d A/C/G TP and 4 mm du TP) 1 ml BR dntp/du TP Mi x (2 mm d A/C/G TP and 4 mm du TP) 5 x 1 m l dntp Sets and dntp/dutp Sets PRODUCT SIZE BR dntp S et ( 4x 1 00 mm solu tio ns) 4 x ml BR dntp S et ( 4x 1 00 mm solu tio ns) 4 x 1 m l BR dntp S et ( 4x 1 00 mm solu tio ns) 5 x 4 x 0.25 ml BR dntp S et ( 4x 1 00 mm solu tio ns) 20 x 4 x ml BR dntp/du TP S et (4x 100 mm so lution s) 4x 0.25 ml Standard PCR Long-range and high-fidelity PCR cdna synthesis and RT-PCR qpcr Sequencing Labeling Exceptional quality dntps of higher than 99% purity confirmed by HPLC Free from DNA and PCR inhibitors Consistent PCR results due to outstanding dntp stability Aqueous solutions (ph 7.0) of Deoxyribonucleatide Triphosphates (datp/dctp/dgtp/dttp or dutp) individually or mixed at indicated concentrations. biotechrabbit Deoxynucleotide Triphosphates are first -choice nucleotides for all PCR applications, including the most demanding ones like amplification of long targets (up to 40 kb), GC-rich templates, qpcr, cdna synthesis, high-fidelity PCR, DNA labeling, and sequencing. Due to advanced production technology, Deoxyribonucle otide Triphosphates are proven to be of a higher than 99% purity and outstanding stability, ensuring excellent performance and consistent, reliable results. For the maximum flexibility, nucleotides are available in sets and mixes of common concentrations. Functional Assays: Tested in qpcr to meet following specifications: 3.0 > Slope > 3.6 Correlation coefficient: > % > efficiency > 90% No template control: no human DNA contamination Physical assays: DNAse: no activity detected RNAse: no activity detected Phosphatase: No activity detected Purity by HPLC: >99% each component 30 Bulk and custom formulations available Please contact oem@biotechrabbit.com

31 DEOXYRIBONUCLEOTIDE TRIPHOSPHATE CHEMICAL FORMULA AND MOLECULAR WEIGHT STRUCTURAL FORMULA datp 2'-deoxyadenosine 5' - triphosphate C 10H 13N 5O 12P 3Na 3 MW (acid form 491.2) dctp 2'-deoxycytidine 5' - triphosphate C 9H 13N 3O 13P 3Na 3 MW (acid form 467.1) dgtp 2'-deoxyguanosine 5' - triphosphate C 10H 13N 5O 13P 3Na 3 MW (acid form 507.2) dttp 2'-deoxythymidine 5' - triphosphate C 10H 14N 2O 14P 3Na 3 MW (acid form 482.1) dutp 2'-deoxyuridine 5'- triphosphate C 9H 12N 2O 14P 3Na 3 MW (acid form 468.1) info@biotechrabbit.com 31

32 Nucleic Acid Purification Pure nucleic acids are essential starting materials for molecular biology methods: gene-expression analyses require pure RNA and genomics methods require pure genomic DNA. The quality of purified RNA or DNA is critical for successful molecular biology. Of the many manual and automated methods available for RNA and DNA purification, spin-column kits are the most frequently used. These kits provide fast and convenient purification of nucleic acids for all demanding application s. Qualitative nucleic acid purification kits are used for recovering high amounts of pure material and requir e little hands-on time. Universal kits allow purification from different sources, including cells, tissues, blood, bacteria and plants. Source-specific kits typically allow higher yields and purity. biotechrabbit continuously expands its nucleic acid purification portfolio with new universal column-based and source-specific kits. Please refer to the table below for choosing products according to your needs. Nucleic Acid Purification Products Selection Guide Product Applications Starting Material RNA Purification Solution Modified guanidine isothiocyanate/phenol method for the extraction of RNA from cells and tissues Universal kit for total RNA isolation from various sources Simple and efficient RNA isolation from different sources in scalable volumes Tissue samples (200 mg) Eucaryotic cells (5x 10 6 ) Bacterial cells; (5x 10 9 ) DNA Purification Universal kit for both high and low copy number plasmid purification from different volumes of bacterial culture Universal kit for genomic DNA isolation from various eukaryotic starting materials For high-copy plasmids: ml bacterial suspension for low-copy plasmid DNA, (> 5-10 ml bacterial suspension Tissue samples of up to 50 mg Rodent tail (0.5 1 cm) Paraffin samples, Buccal swabs Eucaryotic cells (max ) Plant DNA Kit Plant DNA isolation from various materials Fresh, frozen or dry plant material (max 100 mg) Universal kit for bacterial gdna extraction from gram+ and gram cell cultures Up to bacterial cells Virus DNA/RNA Purification Viral RNA extraction form various eukaryotic samples Simultaneous Viral DNA & RNA extraction from eukaryotic samples Serum, plasma, other cell-free body fluids, supernatants from cell cultures (150 µl) Tissues and biopsies (up to 20 mg), Cell cultures (max ), Swab samples, Paraffin samples Serum, plasma, other cell-free body fluids, supernatants from cell cultures (150 µl) Tissues and biopsies (up to 20 mg), Cell cultures (max ), Swab samples, Paraffin samples DNA Clean-up PCR product clean-up Up to 50 µl PCR Mix Kit PCR product clean-up and DNA extraction from agarose gels TAE or TBE agarose gels (up to 300mg) PCR reaction mixtures (up to 50 µl) DNA extraction from TAE or TBE agarose gels TAE or TBE agarose gels (up to 300mg) 32 Bulk and custom formulations available Please contact oem@biotechrabbit.com

33 NEW UPzol RNA Isolation Solution BR ml So lution BR ml 2 Modified guanidine isothiocyanate/phenol method for the extraction of RNA Simple and efficient RNA isolation from different sources in scalable volumes High yields of high quality RNA in 1 hour Starting material: Tissue samples (100 mg) Monolayer cells Cell suspension (animal, plant, yeast, bacterial cells; 5 x 10 6 ) Time: Approximately 1 hour Yield: Depends on the type and amount of the starting material biotechrabbit efficient isolation of total RNA from animals and cells, bacterial cells, plants and other material in variable amounts. The extraction method is based on a time -saving, one-step liquid phase separation. -phase solution containing phenol and guanidine thiocyanate. After the addition of chloroform and subsequent centrifugation, the homogenate is separated into three phases: Colored organic phase (lower) White interphase (middle) Colorless aqueous phase containing RNA (upper) From this upper aqueous phase, RNA is precipitated with alcohol. RNA extraction requires approximately 1 hour. The -quality and highintegrity RNA which can be used for all downstream applications, including northern analysis, cdna synthesis, RT-PCR, dot-blot hybridization, poly A + selection, in vitro translation, cloning and RN Ase assays. The solution is functionally t ested in RNA extraction. NEW GenUP Total RNA Mini Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents Total RNA isolation from various sources Universal kit for total RNA isolation from various starting materials Fast and simple procedure, DNA removed by prefiltration, No DNAse digestion High yields of pure RNA Starting material: Tissue samples (200 mg) Eukaryotic cells (5 x 106) Bacterial cells (5 x 109) Column binding capacity: 100 µg Time: Approximately minutes Yield: Depends on the type and amount of the starting material biotechrabbit GenUP Total RNA Mini Kit has been specially developed for quick and easy p urification of total RNA from eukaryotic cells, tissues, or biopsies, gram positive and gram negative bacteria, and other sources. After few initial procedures, the RNA is bound to a column, washed and then eluted in a separate tube. The prefiltration step removes DNA and the DN Ase digestion is not required. The purified RNA is ready to be used in all demanding molecular biology applications, including cdna synthesis and northern blotting. The kit was tested by isolation of RNA from tissue samples and subsequent analysis of RNA on Agilent Bio Analyzer. Picture 1. High quality total RNA purified from bacterial pellets with and loaded on the formaldehyde agarose gel. Reproducible results in all 3 samples. info@biotechrabbit.com 33

34 NEW GenUP Plasmid Mini Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents Plasmid DNA isolation from bacterial cultures Universal kit for both high-copy and low-copy number plasmid isolation Fast and simple procedure High yields of pure plasmid DNA for all applications Starting material: ml of bacterial cultures for high copy number plasmids 5-10 ml of bacterial cultures for low copy number plasmids Column binding capacity: 40 µg Time: Approximately 16 minutes Yield: Variable. Approx µg of high copy plasmid DNA is typically purified from 2 ml culture The biotechrabbit GenUP TM Plasmid Mini Kit is designed for a fast and efficient plasmid purification from ml bacterial suspensions in a mini format. After performing an alkaline lysis step and precipitating both chromosomal DNA as well as bacterial proteins, pdna is bound to a Mini Column membrane, washed and then eluted in a low-salt buffer. This typically results in µg plasmid DNA from a 5-10 ml bacterial suspension, depending on a plasmid copy number. The purified plasmid DNA is ready to be used in all demanding molecular biology applications, it can be su bjected to enzymatic reactions, can be sequenced etc. The kit is tested for plasmid DNA purification and subsequent analysis of purified plasmid. Competitor GenUP TM isopropanol Picture 1. Plasmid DNA (pbluescript) purified from bacterial pellets (E. coli JM101) and loaded on agarose gel. From left to right: plasmid yields in µg obtained with competitor kit ( 2x), plasmid yields obtained with GenUP TM Plasmid Mini Kit (2x), Plasmid yields obtained with isopropanol method (2x). GenUP TM Plasmid Mini Kit gives highest yields 34 Bulk and custom formulations available Please contact oem@biotechrabbit.com

35 NEW GenUP gdna Mini Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents DNA isolation from tissues and cells Universal kit for DNA isolation from various starting materials including tissues, cells and paraffin samples Fast and simple procedure > 100 µg gdna column binding capacity High yields of pure DNA Starting material: Eucaryotic cells (up to ) Tissue samples (up to 50 mg) Rodent tail specimens cm in length Paraffin-embedded tissue samples Buccal swabs Column binding capacity: >100 µg Time: Approximately 8 minutes after lysis Yield: Depends on the type and amount of material (up to 65 µg) NEW GenUP Plant DNA Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents Isolation of DNA from plants Simple and efficient procedure for DNA isolation from various plant materials High yields of inhibitor-free DNA Starting material: Fresh, frozen or dried plant biomass (max 100 mg) Column binding capacity: <50 µg Time: Approximately minutes Yield: Depends on the type and amount o f the starting material, up to 25 µg The biotechrabbit GenUP TM gdna Mini Kit is designed for a fast and efficient genomic DNA isolation from various sources and different amounts of starting material, like blood, mammalian tissues, including paraffin -embedded ones, buccal swabs or eukaryotic cell cultures. After a few initial procedures, the genomic DNA is bound to a Mini Column membrane, washed and then eluted in a low-salt buffer. This results in high amounts of pure unshared genomic DNA ready to be used in all demanding molecular biology applications, it can be sub jected to enzymatic reactions, can be sequenced, amplified etc. The kit was tested by genomic DNA purification from tissue samples and subsequent analysis of purified gdna in PCR target amplification Picture 1. GenUP TM gdna Mini Kit provides reproducibly high quality template for PCR. The DNA was isolated from different buccal swab samples with GenUP TM gdna Mini Kit and used to amplify human GAPDH gene fragment under fast cycling conditions. biotechrabbit GenUP Plant DNA Kit has been specially developed for quick and easy purification of plant DNA from a wide variety of plant material like leaves, roots, stems, blossoms, fresh, frozen or dried samples. The kit includes an advanced prefiltration step to get rid of unlysed plant biomass. After few procedures, the DNA is bound to a column, washed and then eluted in a separate tube. The purified DNA is ready to be used in all demanding mol ecular biology applications, including PCR, enzymatic digestions, cloning and other. The kit was tested by plant DNA purification and subsequent analysis of purified DNA in PCR target amplification Picture 1. GenUP TM Plant DNA Kit provides reproducibly high quality Plant DNA. The DNA was isolated from different orcideen species and about 2% of total eluate was loaded on the gel. info@biotechrabbit.com 35

36 NEW GenUP Bacterial gdna Mini Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents Isolation of bacterial gdna from both gram+ and gram - species Universal kit for both gram+ and gram-bacteria DNA extraction Fast and simple procedure, high starting volumes of the bacterial culture High yields of pure DNA for demanding applications Starting material: Gram- bacterial cells (1x 10 9 ) Gram+ bacterial cells (1x 10 9 ) Column binding capacity: <50 µg Time: Approximately 45 minutes Yield: Up to 35 µg, depends on the type and amount of the starting material biotechrabbit GenUP Bacterial gdna Mini Kit has been specially developed for quick and easy purification of bacterial genomic DNA from both gram - and difficult to proceed gram+ bacterial cultures. The combined lysozyme and proteolytic lysis steps allow for efficient cell disruption. After few initial procedures, the RNA is bound to a high capacity column, washed and then eluted in a separate tube. The purified DNA is ready to be used i n all demanding molecular biology applications, including PCR, enzymatic digestions, cloning and other. The kit was tested by isolation of gdna from bacterial cultures and subsequent analysis on Agilent BioAnalyzer. Picture 1. GenUP TM Bacterial gdna Mini Kit provides reproducibly high quality DNA even from difficult to proceed gram+ species. The DNA was isolated from 3 different gram+ bacteria cultures and loaded on the gel. 36 Bulk and custom formulations available Please contact oem@biotechrabbit.com

37 NEW GenUP Virus RNA Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents Viral RNA purification form eukaryotic samples Universal kit for viral RNA isolation from various starting materials Fast and simple procedure Flexible elution volumes High yields of pure RNA Starting material: Tissue samples, biopsies (<20 mg) Eukaryotic cells (5 x 10 6 ) Cell-free body fluids, serum, plasma, cell culture supernatants (150 µl) Swab samples Paraffin embedded tissues Time: Approximately 25 minutes Yield: Depends on the type and amount of the starting material Virus RNA Kit has been specially developed for quick and easy purification of viral ssrna from eukaryotic samples including tissues, plasma, serum, other fluids, tissues imbedded in paraffin, cell cultures and buccal swabs. The unique binding membrane of our spin columns has high capacity and guaranties high yields. The high concentration of the purified RNA can be achieved due to flexible elution volumes. After few initial procedures, the viral RNA is bound to a column, washed and then eluted in a separate tube. The purified RNA is ready to be used in all demanding molecular biology applications, including cdna s ynthesis and northern blotting. The kit was tested by isolation of virus RNA and subsequent qpcr. info@biotechrabbit.com 37

38 NEW GenUP Virus RNA/DNA Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents Simultaneous viral DNA and RNA isolation from various sources. Excellent for unknown viruses. Universal kit for simultaneous RNA and DNA isolation from various starting materials Fast and simple procedure, sample specific protocols High yields of pure RNA and DNA Starting material: Tissue samples, biopsies (<20 mg) Eukaryotic cells (5 x 10 6 ) Cell-free body fluids, serum, plasma, cell culture supernatants (150 µl) Swab samples Paraffin embedded tissues Time: Approximately 25 minutes Yield: Depends on the type and amount of the starting material developed for a simultaneous isolation of viral RNA and DNA by keeping the procedure quick and easy. The kit is especially useful when the origin of viruses is unknow. Viral dsdna and ssrna together can be isolated from eukaryotic samples including tissues, plasma, serum, and other fluids, tissues imbedded in paraffin, cell cultures and buccal swabs. The unique binding membrane of our spin columns has high capacity and guaranties high yields. The high concentration of the purified RNA can be achieved due to flexible elution volumes. After few initial procedures, the viral nucleic acids are bound to a column, washed and then eluted in a separate tube. The purified material is ready to be used in all demanding molecular biology applications, including cdna synthesis, northern blotting, qpcr, and RT-PCR. The kit was tested by isolation of virus RNA and DNA and subsequent qpcr. Picture 1. Isolation of viral RNA (FMD Virus) from 3 different dilutions in serum sample. Virus RNA was detected after TaqMan qpcr. ensitive virus detection compared to Competitor (green) kit. 38 Bulk and custom formulations available Please contact oem@biotechrabbit.com

39 NEW GenUP PCR Cleanup Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR preps Multip le com pon ents PCR cleanup Fast and convenient 2-step PCR cleanup procedure No washings, just binding and elution of DNA. 2x shorter procedure compared to competitors High-purity DNA recovery for all demanding applications Starting material: Up to 50 µl PCR Mix Column binding capacity: >20 µg Time: Approximately 3 minutes Recovery: Up to 95% GenUP PCR Cleanup Kit has been developed for quick and easy cleanup or concentration of PCR fragments from reaction mixtures. DNA is bound to a mini column using a novel buffer and eluted. The washing step is eliminated and a lot of hands -on-time is saved. The procedure takes about 3 minutes instead of 8 minutes required with other kits. The purified DNA is ready to be used in all demanding molecular biology applications, including restriction digestion, ligation and sequencing. The components of the kit were tested by purification of DNA from PCR mixtures and subsequent analysis of the recovered DNA. NEW GenUP PCR/Gel Cleanup Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR preps Multip le com pon ents PCR cleanup and DNA extraction from agarose gels Dual performance kit for both PCR product cleanup and DNA purification from agarose gels Fast and simple procedure High DNA recovery yields up to 95% Purification of long DNA up to 30 kb Startin g materia l: TAE or TBE a garos e gel s (up to 300 mg) PCR mi xtures (up to 50 µ l) Extra ction ti me Approx. 20 m inu tes Approx. 3 mi nutes Bind in g capacit y >2 0 µg DN A >2 0 µg DN A DNA si ze 100 bp 30 kb 60 bp 30 k b GenUP PCR/Gel Cleanup Kit has been specially developed for quick and easy cleanup or concentration of PCR fragments from reaction mixtures as well as for isolation of DNA from both TAE and TBE agarose gels. After few initial procedures, the DNA is bound to a column using a novel buffer, washed and then eluted in a separate tube. Particular in PCR cle anup protocol, the washing step is eliminated and a lot of hands-on-time is saved. The procedure takes about 3 minutes instead of 8 minutes required with other kits. The purified DNA is ready to be used in all demanding molecular biology applications, including enzymatic reactions (e.g., restriction digestion and ligation), sequencing, transfection into mammalian cells, and in vitro transcription. The components of the kit were tested by purification and recovery of DNA fragments from agarose gel and PCR mixtures and subsequent analysis of the recovered DNA on Agilent BioAnalyzer. Recov er y rate % % info@biotechrabbit.com 39

40 NEW GenUP Gel Extraction Kit BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents BR pr eps Multip le com pon ents DNA extraction from TAE and TBE agarose gels Fast and simple DNA extraction from agarose gels High DNA recovery yields up to 95% Purification of long DNA up to 30 kb Excellent purity DNA for demanding applications Startin g mater ial TAE or TBE a garos e gels (up to 300 mg) GenUP Gel Extraction Kit has been specially developed for quick and easy extraction of DNA from both TAE and TBE agarose gels. After few initial procedures, the DNA is bound to a mini column using a novel buffer, washed and then eluted in a separate tube with flexible elution volumes (10-50 µl). The purified DNA is ready to be used in any demanding molecular biology application, including enzymatic reactions (e.g., restriction digestion and ligation), sequencing, transfection into mammalian cells, and in vitro transcription. The components of the kit were tested by purification and recovery of DNA fragments from agarose gel and subsequent analysis of the recovered DNA on Agilent Bio Analyzer. Extra ction ti me Approx. 20 m inu tes Bind in g ca pacit y >2 0 µg DN A DNA si ze 100 bp 30 kb Recov er y rate % Picture 1. Excellent Sequencing results obtained on the DNA fragment purified with GenUP Gel Extraction Kit 40 Bulk and custom formulations available Please contact oem@biotechrabbit.com

41 DNA Electrophoresis Nondenaturing agarose or polyacrylamide gel electrophoresis provides a fast way to determine the size of DNA fragments and to quantify the DNA band. During electrophoresis, the negatively charged DNA migrates towards the positively charged electrode. As longer the DNA fragments travel slower within the gel than small fragments, the DNA is separated according to size. For successful electrophoresis, it is important to have a high-quality gel and to prepare the DNA sample for electrophoresis correctly. The use of biotechrabbit DNA Loading Dye, 6x for preparation of DNA samples prior to loading ensures the sample settles into the gel well. Tracking dyes allow migration through the gel to be monitored. After electrophoresis, the DNA is typically stained with intercalating dye that allows DNA visualization directly in the gel under UV light. The detection limit for ethidium bromide is 1 10 ng DNA and for SYBR Green I or similar dyes is 60 pg. For DNA sizing and quantification, the band of interest is compared with the bands of the DNA ladder loaded in the same run. The quality of the ladders used determines the precision and accuracy of these calculations. We recommend biotechrabbit 1 kb DNA Ladder, 50 bp DNA Ladder and 100 bp DNA Ladder for the best results: Product Applications Main features 1 kb DNA Ladder 100 bp DNA Ladder 50 bp DNA Ladder Approximate DNA sizing and quantification within the gel Approximate DNA sizing and quantification within the gel Approximate DNA sizing and quantification within the gel bp range Room-temperature stable Ready to use, sharp bands bp range Room-temperature stable Ready to use, sharp bands bp range Ready to use, sharp bands DNA Loading Dye, 6x Preparation of DNA samples for loading into the gel Free from nucleases, ensures excellent electrophoresis results info@biotechrabbit.com 41

42 DNA Electrophoresis Ladders and Loading Dyes PRODUCT SIZE BR BR NE W BR BR kb D NA Ladd er, rt u 100 bp DNA Ladd er, rt u 50 b p D NA Ladd er, rt u DNA Load ing Dye, 6 x 100 lanes (5 µ l) 100 lanes (5µ l) 100 lanes (5µ l) 500 µ l 1 kb DNA Lad der, rtu 1ml D NA L oadin g Dye, 6 x 500 µ l 100 bp DNA La dde r, rtu 1ml D NA L oadin g Dye, 6 x 500 µ l 50 bp DNA Lad der, rtu 1ml D NA L oadin g Dye, 6 x 5 x 1 m l 5 x 1 ml DNA L oadin g Dye, 6 x biotechrabbit DNA electrophoresis ladders are mixtures of exceptionally purified DNA fragments created either by PCR or by digesting proprietary plasmids with restriction enzymes. Ladders are ready to use, and suitable not only for DNA sizing but also for approximate DNA quantification in gels. For convenience, ladders have increased intensity reference bands and indicated DNA amount i n nanograms for every band. Every ready-to-use ladder is supplied with the nuclease -free Loading Dye Solution, which ensures optimal migration and quantification of your DNA probes. DNA sizing and approximate quantification on agarose gels Ready-to-use DNA ladders ideal for DNA sizing and quantification on the gel Pure and stable: all ladders can be stored at +4 C, some even at room-temperature Every ladder supplied with 6x Loading Dye for sample DNA STORAGE AND STABILITY Exceptionally stable 1 kb and 100 bp DNA Ladders can be conveniently stored under following conditions: At 25 C for 6 months At 4 C for 12months At -20 C for 24 months The 50 bp DNA Ladder can be stored: At 4 C for 12months At -20 C for 24 months Prod uct Com posit ion DNA Conc. 1 kb DN A Ladd er, r tu 100 bp DN A Ladd er, r tu 50 bp DNA Ladd er, r tu DNA Loadi ng Dye, 6x DNA i n 10 mm Tris -HCl (ph 8.0) and 10 mm E DTA, glycerol a nd tra cki ng d yes DNA i n 10 mm Tris -HCl (ph 8.0) and 10 mm E DTA, glycerol a nd tra cki ng d yes DNA i n 10 mm Tris -HCl (ph 8.0) and 10 mm E DTA, glycerol a nd tra cki ng d ye 10 mm Tri s -H Cl (ph 8.0) and 60 mm E DTA, gl ycerol a nd track ing d yes Tested in gel electrophoresis. Not appli cabl e Ele ctro phore sis tracking d yes Bromo phenol blu e Xylene cyanol FF Orang e G Xylene cyanol FF Orang e G Bromo phenol blu e Orang e G Xylene cyanol FF 1 kb DNA Ladder, rtu 100 bp DNA Ladder, rtu NE W 50 bp DNA Ladder, rtu DNA Loading Dye, 6x Range : bp Refer ence : and bp Nu mber of bands : 1 3 Range : bp Refer ence : 5 00 and b p Nu mber of bands : 1 2 Range : bp Refer ence : 200 and 50 0 b p Nu mber of bands : 1 7 Appro ximat e mi grati on o f tracking D yes in 1 % TA E agarose ge l, am bient light. X yl e ne c ya no l F F ~ 4 kb Bro m o p he no l b l ue ~400 b p Ora nge G~ > 5 0 b p 42 Bulk and custom formulations available Please contact oem@biotechrabbit.com

43 Ultrapure Enzymes for Molecular Biology Product Applications Main features phi 29 DNA Polymerase Rolling circle amplification (RCA) Multiple displacement amplification (MDA) Whole genome amplification (WGA) Protein-primed DNA amplification RNA-primed DNA amplification Huge DNA yields, accurate amplification Highest processivity DNA polymerase with exceptionally strong strand -displacement activity StranDisplace Thermostable DNA Polymerase Loop-mediated isothermal amplification (LAMP) Sequencing Exceptionally pure thermophilic DNA polymerase for demanding applications Strong strand-displacement activity T4 DNA Ligase, Rapid Blunt and sticky end DNA ligation for cloning Joining of linkers or adaptors to double-stranded DNA Self-circularization of linear DNA Exceptionally pure, high concentration T4 DNA Ligase for demanding applications Rapid ligation Supplied with two buffers for standard and fast ligation protocols Uracil DNA Glycosylase PCR carry-over contamination control Exceptionally pure Uracil demanding applications DNA Glycosylase for 43

44 phi 29 DNA Polymerase, 10 U/µl BR BR units 1000 units 25 µ l p hi 29 DNA Pol ym erase, 1 0 U /µl 1 ml 10x p hi 2 9 R ea ction Buffer 100 µl p hi 29 DN A Pol ymerase, 10 U/µl 1 ml 10x p hi 2 9 R ea ction Buffer Rolling circle amplification (RCA) [ 5] Multiple displacement amplification (MDA) [6] Whole genome ampli fication (WGA) [7] Protein-primed DNA amplification [ 8] RNA-primed DNA amplification [9] Huge DNA yields, accurate amplification Highest processivity DNA polymerase with exceptionally strong strand -displacement activity Exceptionally pure phi 29 DNA Polymerase for demanding applications phi 29 DNA Polymerase, 10 U/µl in Storage Buffer: 10 mm Tris-HCl, 100 mm KCl, 0.1 mm EDTA, 1 mm DTT, 0.5% Tween-20, 0.5% NP-40, 50% glycerol, ph 7.4 at 25 C 10x phi29 Reaction Buffer: 500 mm Tris-HCl, 100 mm (NH 4) 2SO4, 40 mm DTT, 100 mm MgCl2, ph 7.5 at 25 C biotechrabbit phi 29 DNA Polymerase is an exceptionally pure DNA polymerase for demanding applications. The enzyme is a highly processive DNA polymerase (up to 70,000 base insertions per binding event) with a powerful strand - displacement activity [1] and a 3' 5' proofreading exonuclease function [2]. phi29 DNA Polymerase proofreading activity acts preferentially on single-stranded DNA or RNA. Therefore, to avoid primer degradation during the DNA synthesis, 3'-modified (protected) primers are highly recommended [3]. phi29 DNA Polymerase is responsible for the replication of the Bacillus subtilis phage phi 29 [4]. The enzyme is purified from a recombinant E. coli strain carrying the phi29 DNA Polyme rase gene from bacteriophage phi29. REFERENCES: 1. Blan co, L., et a l. (19 89) J. Bio l. Ch em., 264, Garmen dia, C., e t a l. (199 2) J. Bio l. C hem., 267, Skerra, A., (199 2) Nuc le ic Aci ds Res., 20, Blan co, L., an d Salas, M., (1984) Pr o c. Nat l. A cad. S ci. U SA, 8 1, Liza rdi, P. M., et al. (1998) Nat. Gen et., 19, Dean, F.B., et a l. ( 200 2) Pro c. Natl. Acad. Sc i. USA, 99, Sim me l, F. C., et al. (2005) Nan o L ett., 5, Blan co, L., et a l. (199 4) Pr oc. Nat l. A cad. Sc i. USA, 91, Lagunavic ius, A., et al. (2009 ) R NA, 1 5, UNIT DEFINITION One unit is defined as the amount of polymerase required to convert 0.5 pmol of dttp into acid insoluble material in 10 minutes at 30 C. Single-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rdna Contamination Test 44 Bulk and custom formulations available Please contact oem@biotechrabbit.com

45 StranDisplace Thermostable DNA Polymerase, 40 U/µl BR units 200 µ l S trandis place Thermos tabl e DNA Pol ym erase, 4 0 U /µl 8 x 1 ml 10x Po lymerase R ea cti on Buff er Loop-mediated isothermal amplification (LAMP) [ 2, 3] Sequencing [4] Exceptionally pure thermophilic DNA polymerase for demanding applications Strong strand-displacement activity StranDisplace Thermostable DNA Polymerase, 40 U/µl in Storage Buffer: 10 mm Tris-HCl, 50 mm KCl, 1.0 mm DTT, 0.1 mm EDTA, 0.1% Triton X-1, 50% glycerol, ph 7.5 at 25 C 10x Polymerase Reaction Buffer: 200 mm Tris-HCl, 100 mm (NH 4) 2SO4,100 mm KCl, 20 mm MgSO4, 1.0% Triton X-100, ph 8.8 at 25 C biotechrabbit StranDisplace Thermophilic DNA Polymerase is an exceptionally pure DNA polymerase for applications in which strong strand-displacement activity at elevated temperatures is required. The StranDisplace Thermophilic DNA Polymerase is a thermophilic DNA polymerase with a strong strand -displacement activity and is deficient in both proofreading (3 ' 5') and nicktranslation (5' 3') nuclease activities. The protein was originally characterized and its crystal st ructure solved by Lorena Beese [ 1]. StranDisplace Thermophilic DNA Polymerase can be used for the same applications as Bst DNA Polymerase Large Fragment and Bsm DNA Polymerase. The enzyme is purified from a recombinant E. coli strain carrying the engineered Bacillus stearothermophilus DNA polymerase gene fragment. REFERENCES: 1. Kie fer, J.R., et a l. (1997) Stru ctur e, 5, Noto mi, T., et a l. ( 20 00) Nu cleic Ac i ds Re s., 28, e6 3 E pub. 3. Imai, M., et a l. ( 200 7) J. Vir ol. Meth o ds, 14 1, Mead, D.A., et al. (1991) Biote chni qu es, 11, UNIT DEFINITION One unit is defined as the amount of polymerase required to convert 10 nmol of dntps into acid insoluble material in 30 minutes at 65 C. Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rdna Contamination Test info@biotechrabbit.com 45

46 T4 DNA Ligase, Rapid, 600 U/µl BR BR units units 100 µl T4 DN A Liga s e, Rapid, 600 U /µl 1 ml 2 x Ra pid Liga tion Buffer 1 ml 10x Li gation Buffer 300 µl T4 DNA Liga se, Rapid, 600 U /µl 3 x 1 ml 2 x Rap id Ligation Buffer 3 x 1 ml 10x Li gati on Buff er Blunt- and sticky-end DNA ligation for cloning Joining of linkers or adaptors to double-stranded DNA Self-circularization of linear DNA Exceptionally pure high-concentration T4 DNA Ligase for demanding applications Rapid ligation Supplied with two buffers for standard and fast ligation protocols T4 DNA Ligase, Rapid, 600 U/µl in Storage Buffer: 10 mm Tris-HCl, 50 mm KCl, 1 mm DTT, 0.1 mm EDTA, 50% glycerol, ph 7.5 at 25 C 10x Rapid Ligation Buffer: 132 mm Tris-HCI, 20 mm MgCl2, 2 mm DTT, 2 mm ATP, 15% PEG 6000, ph 7.6 at 25 C 10x Ligation Buffer: 500 mm Tris-HCI, 100 mm MgCl 2, 50 mm DTT, 10 mm ATP, ph 7.6 at 25 C biotechrabbit T4 DNA Ligase, Rapid, is an exceptionally pure, highly concentrated ligase for applications in which high concentrations of the enzyme are required. It is especially recommended for fast ligations. T4 DNA Ligase, Rapid, is supplied with two buffers : the common 10x Ligation Buffer for typical 1 hour or overnight ligations and the 2x Rapid Ligation Buffer contai ning PEG for fast 5 10 minutes ligations or ligation of low-concentration or blunt -end DNA. The 10x Ligation Buffer does not contain PEG and is compatible with standard ligation protocols which do not specify the use of a rapid/fast/quick format buffer. The T4 DNA Ligase, Rapid, in combination with the 2X Rapid Ligation buffer greatly stimulates the rate and efficiency of bluntend ligation. Long incubations (>10 minutes) are not recommended as they reduce the transformation efficiency of ligation product s. T4 DNA Ligase catalyzes the formation of a phosphodiester bond between the terminal 5 ' phosphate and the 3' hydroxyl groups of duplex DNA or RNA. The enzyme efficiently joins blunt and cohesive ends and repairs single -stranded nicks in duplex DNA, RNA or DNA/RNA hybrids [1]. The enzyme is purified from a recombinant E. coli strain carrying the cloned T4 DNA Ligase gene. REFERENCES: 1. Engler, M.J., and Ri chardson, C. C., ( 1982) P.D. Bo ye r (E d.), The Enzym es, V ol. 5, pp. 3. San D ie go: A cade mic Pr ess. UNIT DEFINITION One unit is defined as the amount of T4 DNA Ligase required to join 50% of 100 ng of DNA fragments with cohesive termini in 50 µl 1x T4 DNA Ligase Buffer following 30 minute incubation at 23 C. Notes: One biotechrabbit T4 DNA Ligase cohesive end unit is equivalent to approximately 3 cohesive end units as measured with a Lambda-Hind III DNA fragment substrate in 1 x T4 DNA Ligase reaction buffer. One Weiss Unit is approximately equivalent to 22 biotechrabbit cohesive end units. Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rdna Contamination Test 46 Bulk and custom formulations available Please contact oem@biotechrabbit.com

47 Uracil DNA Glycosylase, 2 U/µl BR BR U 1000 U 100 µl Uracil DN A Gl ycos ylase, 2 U /µl 1 ml 10x U DG R eactio n Buff er 500 µl Uracil DN A Gl yco s ylase, 2 U /µl 2 x 1 ml 1 0x UDG R ea ction Buffer PCR carry-over contamination control [1] Site-directed mutagenesis [2] Glycosylase mediated S NP detection (GMPD) [ 3] Exceptionally pure Uracil DNA Glycosylase for demanding applications Uracil DNA Glycosylase, 2 U/µl in Storage Buffer: 10 mm Tris-HCl, 50 mm NaCl, 1 mm DTT, 0.1 mm EDTA, 50% glycerol, ph 7.5 at 25 C 10x UDG Reaction Buffer: 200 mm Tris-HCI, 10 mm DTT, 10 mm EDTA, ph 8.0 at 25 C biotechrabbit Uracil DNA Glycosylase is an exceptionally pure enzyme which catalyzes the hydrolysis of the N - glycosylic bond between uracil and sugar, leaving an abasic site in uracil-containing single or double -stranded DNA. The enzyme shows no measurable activity on short oligonucleotides (<6 bases), or RNA substrates. The enzyme is purified from a recombinant E. coli strain carrying the Uracil DNA Glycosylase gene from E. coli K-12. REFERENCES: 1. Lon go, M.C., et al. (1990) Gene, 93, Kun ke l, T.A., ( 1985) Pro c. Natl. Aca d. S ci. USA, 8 2, Vau ghan, P., an d Mc Carth y, T.V., ( 19 98) N uc lei c A cids R es., 26, UNIT DEFINITION One unit is defined as the amount of enzyme that catalyzes the release of 1.8 nmol of uracil in 30 minutes from double - stranded, tritiated, uracil-containing DNA at 37 C in 1x UDG Reaction Buffer. Note: Two units of Uracil DNA Glycosylase degrade about 0.2 µg Uracil containing DNA in 10 minutes at 37 o C. DNA degraded in this manner cannot be PCR amplified. Enzyme is active in a broad ph range and in most PCR buffers, with or without divalent cations. However, it is inhibited by high ionic strength (>200 mm). Single-Stranded Exonuclease Activity Double-Stranded Exonuclease Activity Double-Stranded Endonuclease Activity E. coli 16S rdna Contamination Test info@biotechrabbit.com 47

48 Protein Electrophoresis Denaturing SDS polyacrylamide -gel electrophoresis is the most common technique for protein analysis. Proteins are completely denatured in the presence of SDS, DTT and other reagents and loaded under denaturing conditions in Tris-glycine SDS electrophoresis buffer. Subsequent electrophoresis allows protein size to be determined according to electrophoretic mobility by comparing the sample size to similar sized bands of the protein ladder loaded on the same gel. High-quality prestained protein ladders allow approximate molecular weight determination for denatured proteins as well as monitoring of the electrophoresis process and the efficiency of the western transfer. Pr estained protein ladders, however, are not used for precise protein molecular weight determination due to dyes bound to ladder proteins. For approximate protein molecular-weight determination and monitoring of the western transfer efficiency, we recommend biotechrabbit the TriColor Broad Protein Ladder and TriColor Protein Ladder prestained protein ladders: Product Range Number of bands Applications Features TriColor Broad Protein Ladder TriColor Protein Ladder kda kda Approximate protein sizing on SDS PAGE and western blots. Monitoring protein gel electrophoresis and western transfers Ready-to-use, prestained protein ladders for approximate protein sizing, monitoring of electrophoresis and western transfer. Pure and stable: retain sharp bands after 3 months storage at +4 o C 48 Bulk and custom formulations available Please contact oem@biotechrabbit.com

49 Protein Electrophoresis Ladders PRODUCT SIZE BR BR TriCol or Broad Protei n Lad d er, r tu ( kda) TriCol or Protei n Ladder, rtu ( kda) 100 mi ni-gel appli catio ns (5µl /load) 100 mi ni-gel appli catio ns (5µl /load) 500 µl Tri Color Broad Pro tein Ladd er, r tu 500 µl Tri Color Protei n Lad d er, r tu Approximate protein sizing on SDS PAGE and western blots Monitoring of protein gel electrophoresis and western transfer Ready-to-use prestained protein ladders for approximate protein sizing, monitoring of electrophoresis and western transfer Pure and stable: sharp bands after 3 months storage at +4 o C Prod uct Bu ffer c omp ositi on Protein concent ration biotechrabbit TriColor Protein Ladders are perfect tools for an approximate MW estimation of proteins on denaturing polyacrylamide gels, monitoring protein separation during electrophoresis and verifying of western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose). The ladders have sharp protein bands in three colors with easily recognizable colored reference bands. Ladders are available in two versions: TriColor Broad Protein Ladder covering kda range and TriColor Protein Ladder covering kda range. Both are ready for immediate use, are stable and can be stored either for two weeks at room temperature or even for 3 months at +4 o C that eliminates the need to thaw them every time before the load. STORAGE AND STABILITY Exceptionally stable Protein Ladders can be conveniently stored under following conditions: At 25 C for 2 weeks At 4 C for 3months At -20 C for 12 months TriCol or Broad Protei n Lad d er, r tu TriCol or Protei n Ladd er, r tu 20 mm Tris -p ho sp hate ( ph 7.5 a t 25 C), 2% SDS, 1 0 mm Di thio threi tol, 3.6 M urea, 15% gl ycerol 20 mm Tris -p ho sp hate ( ph 7.5 a t 25 C), 2% SDS, 1 mm 2-m erca pto ethanol, 3.6 M ur ea, 15% gl ycerol ~ 0.1~ 0.4 mg /ml, each pro tein ~ 0.2~ 0.4 mg /ml, each pro tein Tested in SDS-Polyacrylamide gel electrophoresis and western transfer. TriColor Broad Protein Ladder ( kda), rtu TriColor Protein Ladder ( kda), rtu Range: kda Reference bands: 25 and 75 kda Number of bands: 13 Range: kda Reference bands: 25 and 75 kda Number of bands: 10 Guide for molecular weight estimation (kda) Migration patterns in different electrophoresis conditions. The apparent molecular weight of each protein (kda) was determined by calibrating against unstained proteins under same conditions. info@biotechrabbit.com 49

50 Protein Purification Media biotechrabbit Agarose 40 Resin is produced from agarose using a proprietary cross -linking method that results in a highly porous and physically stable matrix with a narrow particle size distribution of approximately 40 microns. Agarose-based matrices are successfully used in biotechnology research and in the industrial purification of proteins. Agarose is proven to be exceptionally compatible with natural biomolecules such as proteins, DNA and carbohydrates. Due to the hydrophilic nature, the resin exhibits minimal nonspecific interaction. Unlike matrices from synthetic polymers, agarose does not have micro pores that can contribute to local ph variations in the microenvironment in the column that can lead to distorted separation. Picture 1. Agarose net scheme Agarose is quite inert with respect to soluble molecules. Ligands attached to agarose function -specific background, ensuring low risk of artifacts and easier cleaning. These low levels of non-specific interaction are due to the following features: Very low matrix content: agarose media has typically only 6% matrix content per volume. Synthetic matrices have typically 30% matrix content and correspondingly higher risk of interaction. Very high hydrophilicity: most soluble molecules will not interact with hydrophilic surfaces. Synthetic media are more or less hydrophobic even when they have been modified to increase hydrophilicity. Some synthetic media, such as divinylbenzene, are more hydrophobic; others, such as methacrylate, are less. Very high porosity: pores in synthetic media increase the surface area available for interaction. Agarose does not have micropores, and, accordingly non-specific interaction is limited. In comparison, cellulose, which is a natural, hydrophilic material, has higher risk of non-specific interaction than agarose. Agarose exhibits extreme chemical and physical stability, which is advantageous for process flexibility, economics and process performance: Properly cross-linked agarose has a stability range of ph Many synthetic resins are not stable at high ph. Agarose maintains a nearly constant volume in different solvents and conditions (i.e., does not swell or shrink). Thus, it can be used for extended periods in packed columns or filters without affecting functionality. The surface of an agarose bead is very smooth and the be ad exhibits some elasticity. Even when used for extended periods in columns and filters or when handled extensively during packing and other operations, it will not break up into particles can block the flow. (In comparison, cellulose typically fragments easily on handling ). Agarose is known to have extremely low levels of leach ing and that leachates are carbohydrates having little or no toxicity. Many synthetic media have been used for many years extensively at the laboratory scale but, due to tiny amounts of leakage of charged groups, use cannot be scaled up. Choice of Agarose 40 Media for different applications: Immobilized metal affinity chromatography (IMAC) media and His-tagged protein purification media (IMAC based) IDA/high Agarose 40 Resin IDA/low Agarose 40 Resin TREN/high Agarose 40 Resin TREN/low Agarose 40 Resin Ni Agarose 40 Resin Ion-exchange chromatography protein purification media (IEX) Q Agarose 40 Resin S Agarose 40 Resin Size-exclusion chromatography protein purification media (SEC) SEC Agarose 40 Resin SEC 100 Agarose 40 Resin SEC Agarose 40 Resin 50 Bulk and custom formulations available Please contact oem@biotechrabbit.com

51 His-Tagged Protein Purification Media PRODUCT SIZE BR Ni A garos e 40 R esi n 25 ml* BR Ni A garos e 40 R esi n 15 0 ml* * Conc entrated s lurr y: eq ua l to d ou ble vo lu mes of 50 % s lu rries fr om other s up pli ers. Our pa cka ge si ze corr elat es t o f inal packed co lu mn v olum e. Ni Agarose 40 Resin is developed for capture of His - tagged proteins. Typical binding and wash buffer is 20 mm sodium phosphate, ph 7.4, with 0.5 M NaCl. To reduce unspecific binding, mm imidazole is added to the buffer. Proteins are typically eluted with mm imidazole, although elution conditions must also be determined for each protein. The concentrated suspension provides gel volumes that are twice as large, allowing twice as many applications, than 50% slurries from other suppliers. High-throughput agarose media for capture of Histagged proteins Very high capacity 60 mg/ml High purity with high-flow characteristics COMPOSITION Ni Agarose 40 Resin is supplied in an aqueous suspension containing 22% ethanol as preservative and is ready for use after washing. NOTE: Ni Agarose 40 Resin is provided as a concentrated slurry and can be used at half the volume of 50% slurries from other suppliers. Ni 2+ is the preferred metal ion for purification of His -tagged proteins. Immobilized metal ion affinity chromatography (IMAC) is based on interaction between chelated transition metal ions and side-chains of amino acids (mainly histidine) on proteins. Ni Agarose 40 Resin is produced from agarose using a proprietary cross-linking method that results in a highly porous and physically stable agarose matrix. Ni Agarose 40 Resin for immobilized metal affinity chromatography is activated and a chel ator is coupled according to the Bromohydrin method. This method gives rise to a spacer arm between the agarose backbone and the attached chelator. Ni 2+ ions are already preloaded and the product is ready for use once packed into a column. Agarose media are generally easy to pack. biotechrabbit concentrated Ni agarose 40 Resin is an economical, high-quality product providing a double volume of the packed gel or two times more applications compared to 50% Ni agarose slurries from other suppliers. Our package size correlates to final packed column volume. Tested in standard protein purification scheme. CHARACTERISTICS Agaros e c ontent % Metal i on ca pacit y µeqv Ni 2+ /ml Partic le si ze µ m Protein ca pacit y Max flow rat e at 20 c m bed h ei ght an d 5 bar >6 0 mg /m l >5 00 cm /h ph stabi lit y ph 2 14 So lvent stabi lit y 100% m ethanol 100% ethanol 8 M urea 6 M guani di ne hydrochl orid e 30% a ceto ni tril e 70% for mi c a cid 30% trifluoroa ceti c a ci d info@biotechrabbit.com 51

52 Immobilized Metal Affinity Chromatography Media (IMAC) PRODUCT SIZE BR IDA /hig h Agaro se 4 0 Resi n 25 ml* BR IDA /hig h Agaro se 4 0 Resi n 15 0 ml* BR IDA /lo w Agaro se 4 0 Resi n 25 ml* BR IDA / low Agaros e 4 0 R es in 15 0 ml* BR TREN/hi g h Agaros e 4 0 R esin 25 ml* BR TREN /hig h A garos e 40 R esi n 15 0 ml* BR TREN /lo w A garos e 40 R esi n 25 ml* BR TREN / lo w Agaros e 4 0 R esin 15 0 ml* * Conc entrated s lurr y: eq ua l to d ou ble vo lu mes of 50 % s lu rries fr om other s up pli ers. Our pa cka ge si ze corr elat es t o f inal packed co lu mn v olum e. To help you find the best match for your protein, biotechrabbit offers four different IMAC chemistries. We recommend starting with IDA/high, as this has the best capacity and works well for most proteins. Proteins that are difficult to recover or exhibit lower activity after purification may be binding too strongly. I n these cases, try IDA/low. For further optimization, we recommend trying TREN/high then TREN/low. High-throughput agarose media for immobilized metal affinity chromatography (IMAC) Choice of IMAC chemistry to fit variety of proteins High flow characteristics Agarose 40 Resin IMAC media are produced from agarose using a proprietary cross -linking method that results in a highly porous and physically stable agarose matrix. Agarose 40 Resin IDA and TREN series of immobilized metal affinity chromatography (IMAC) gels are activated and coupled according to the Bromohydrin method. This method provides a spacer arm between the agarose backbone and the attached chelator. Ligand density has been shown to have a great impact on separation. In some case s, a high-capacity gel is needed while in other cases a low-capacity gel is sufficient [1]. Therefore, Agarose 40 Resin IDA and TREN media are available in two degrees of substitution (low and high) to allow maximum flexibility. REFERENCES: 1. Enzym ology 4, 4 13 (199 2) CHELATING GROUPS Chelators attached to Agarose 40 Resin are iminodiacetic acid (IDA, Picture 1), and the new chelator tris(2 - ethylaminoethyl)amine (TREN, Picture 2). COMPOSITION Media are supplied in aqueous suspensions containing 22% ethanol as preservative and are ready for use after washing. CHARACTERISTICS Agaros e 40 R esin IDA TREN Agaros e c ontent, % Chelat in g gro up IDA TREN Metal i on ca pacit y, µeqv N i 2+ /ml I DA lo w I DA hig h Partic le s Max flow rat e at 20 c m bed h ei ght an d 5 bar, cm/h >5 00 >5 00 ph stabi lit y ph 2 14 ph 2 14 So lvent stabi lit y af ter cou pling the ligan d 100% m ethanol 100% ethanol 8 M urea 6 M guani di ne hydrochl orid e 30% a ceto ni tril e 70% for mi c a cid 30% trifluoroa ceti c a ci d TREN lo w TREN hig h Picture 1. Iminodiacetic acid (IDA) Picture 2. Tris(2-ethylaminoethyl)amine (TREN) Tested in standard protein purification scheme. 52 Bulk and custom formulations available Please contact oem@biotechrabbit.com

53 Ion-Exchange Chromatography Protein Purification Media (IEX) PRODUCT SIZE BR Q Agaros e 4 0 R esin 25 ml BR Q Agaros e 4 0 R esin 200 ml BR S A garos e 40 R esi n 25 ml BR S A garos e 40 R esi n 200 ml * Conc entrated s lurr y: eq ua l to d ou ble vo lu mes of 50 % s lu rries fr om other s up pli ers. Our pa cka ge si ze corr elat es t o f inal packed co lu mn v olum e. Q and S Agarose 40 Resin ion exchange media are designed for high-throughput protein separation under a variety of conditions. The high resolution that can be obtained at high protein loading and high flow rates makes this resin ideal for process applications. The chemical stability allows cleaning-in-place (CIP) protocols using sodium hydroxide to be developed easily. Ion-exchange media for preparative- and bioprocess-scale purification of proteins High throughput and high resolution Reliable and reproducible, easy scale -up High chemical stability for easy c leaning in place COMPOSITION Media are supplied in aqueous suspensions containing 22% ethanol as preservative and are ready for use after washing. CHARACTERISTICS Agaros e 40 R esin Q S Agaros e c ontent, % Protein xa pacit y, m g/ ml BSA, 13 0 IgG, 70 Ionic gro up Quartenar y a min e Sulphoni c a ci d Ionic ca pacit y, mm ol/m l Partic le s ph stabi lit y ph 1 14 ph 1 14 So lvent stabi lit y 100% m ethanol 100% ethanol 8 M urea 6 M guani di ne hydrochl orid e 30% a ceto ni tril e 70% for mi c a cid 30% trifluoroa ceti c a ci d Separation media based on agarose are well known for excellent selectivity. Q and S Agarose 40 Resin have a high selectivity, ensuring protein peaks are well separated with greater distance from each other than comparable products made from synthetic polymers. Thus, these media have the capacity to separate proteins well even when loading large amounts of protein. Resolution is the combined effect of selectivity (distance between peaks) and efficiency (peak width, depending on particle size). Q and S Agarose 40 Resin ion-exchange media are produced from agarose using a proprietary cross -linking method that results in a highly porous and physically stable agarose matrix. The particle size of 40 mm, with a very narrow particle size distribution, in combination with the proprietary cross -linking results in columns packed with very high efficiency and excellent flow characteristics that are well suited to demanding bioprocess applications. Q AGAROSE 40 RESIN COMPARISON WITH OTHER MEDIA Med ia Agaros e content, % Ionic gro up Ion capac ity, mmo l/ m l Partic le si ze, Max lin ear flow rate, cm/h Q Agaros e 4 0 Resi n Leadin g 90 µm media >6 >6 Quarternary am in e Quarternary amin e Leadin g 34 µm m edia Quarternary am in e (aver. 9 0) 34 >5 00 (20 cm bed heig ht/5 bar) ( 15 cm bed heig ht/1 bar) ph stabi lit y ph 1 14 ph 2 14 ph 1 14 <1 50 ( 10 cm b ed heig ht/3 bar) Tested in standard protein purification scheme. Picture 1. Wider Binding Dynamics of Q Agarose 40 Resin info@biotechrabbit.com 53

54 Size-Exclusion Chromatography Protein Purification Media (SEC) PRODUCT SIZE BR SEC Agaros e 4 0 R esin 300 ml BR SEC 100 A garos e 40 R esi n 300 ml BR SEC Agaros e 4 0 R esin 300 ml * Conc entrated s lurr y: eq ua l to d ou ble vo lu mes of 50 % s lu rries fr om other s up pli ers. Our pa cka ge si ze corr elat es t o f inal packed co lu mn v olum e. SEC Agarose 40 Resin gel-filtration media is designed for highperformance protein separation under a variety of conditions. The high resolution that can be obtained makes it ideal for both preparative work and process-scale separation of proteins. High-performance size-exclusion chromatography media for preparative- and process-scale separation of proteins Excellent resolution Robust separation results can be achieved for a wide range of proteins and conditions Chemically stable for cleaning in place COMPOSITION Ni Agarose 40 Resin is supplied in aqueous suspensions containing 22% ethanol as preservative and is ready for use after washing. CHARACTERISTICS SEC Agarose 40 Resin size-exclusion chromatography or gelfiltration media has a high selectivity, ensuring that the protein peaks are well separated with greater distance from each other than comparable products made from synthetic polymers. Thus, the media has the capacity to separate proteins well, even when loading large amounts of protein. All SEC Agarose 40 Resin media are produced from agarose using a proprietary cross -linking method that results in a highly porous and physically stable agarose matrix. The narrow particle size distribution of approximately 40 mm in combination with the proprietary cross-linking results in a media that is easy to pack in columns with very high efficiency and good flow characteristics. Tested in standard protein purification scheme. Agaros e 40 R esin SEC SEC 100 SEC 10,000 Agaros e c ontent, % Exc lusion limit, kda >1 0,000 Separat ion range, Very larg e kda mol ecu les Flow rat e in a 8 x 30 0 mm co lu mn; cm/ min Partic le s ph stabi lit y ph 1 14 ph 1 14 ph 1 14 So lvent stabi lit y after cou plin g the li gand 100% m ethanol, 100% etha nol, 8 M urea, 6 M guanid in e hydro chlorid e, 30% a ceto ni trile, 70% formic acid, 3 0% trifluoroa ceti c a ci d Picture 2. Flow/pressure plot. SEC Agarose 40 Resin, Column 0.8 x 30 cm Picture 1. K av curve for some common proteins. The dimer of thyroglobulin elutes in V o and the other proteins nicely follow the theoretical K av relation 54 Bulk and custom formulations available Please contact oem@biotechrabbit.com

55 NOTES biotechrabbit GmbH makes every effort to include accurate and up -to-date information within this publication; however it is possible that omissions or errors might have occurred. Changes of publication can be made at any time without notice. All mentioned trademarks are protected by law. All biotechrabbit products are designed for use for research purposes only (RUO); they are not recommended or intended for diagnosis or treatment of diseases in humans or animals. biotechrabbit products are to be used only by trained laboratory personnel who are familiar with potential hazards and in laboratories equipped to perform life science res earch. biotechrabbit products may contain chemicals which are harmful if misused. Due care should be taken with all products to avoid direct contact with components that are potentially harmful. are registered Trademarks of Applera Corporation or its subsidiaries. are registered Trademarks of Life Technologies. Bioregistered Trademarks of Bio -Rad Laboratories, Inc. Cepheid, SmartCycler are registered Tradem arks of Cepheid Corp. Illumina are registered Trademarks of Illumina Inc. Eppendorf, Mastercycler, REALPLEX are registered Trademarks of Eppendorf AG. LightCycler, TaqMan are registered Trademarks of Roche Group. Mx3000P, Mx3005P, Mx4000, St ratagene are registered Trademarks of Stratagene. Omni Klentaq is registered Trademark of DNA Polymerase Technology, Inc. QIAGEN, Rotor-Gene are registered Trademarks of QIAGEN GmbH. Quantica is registered Trademark of Jonathan Redfern. Scorpions is registered Trademark of DxS Limited. SYBR is registered Trademark of Molecular Probes, Inc. Takara is registered Trademark of Takara Shuzo Co. Ltd. Techne is registered Trademark of Techne Corporation. Triton is registered Trademark of Union Carbide Corp. and 5,436,149. e is sold under license from DNA Polymerase Technology, Inc., U.S. Patent Nos. 7,462, biotechrabbit GmbH, all rights reserved. info@biotechrabbit.com 55

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