PCR Reagents. The Highest Level of PCR Performance. PCR Enzymes RT-PCR Reagents QPCR and QRT-PCR Reagents dntps

Transcription

1 A m p l i f i c a t i o n PCR Reagents The Highest Level of PCR Performance PCR Enzymes RT-PCR Reagents QPCR and QRT-PCR Reagents dntps

2 Stratagene Has Yo High-Fidelity Application General Cloning TA/UA Cloning Long Targets RECOMMENDED page 6 PfuUltra High-Fidelity DNA s page 7 Easy-A High-Fidelity PCR Cloning Enzyme page 10 EXL DNA GOOD page 8 PfuTurbo DNA s page9 PfuTurbo C x Hotstart DNA page 10 Herculase Enhanced DNA s KITS & MASTER MIXES page 6 PfuUltra PCR Master Mix page 8 PfuTurbo PCR Master Mix page 7 Easy-A PCR Master Mix Stratagene PCR Enzymes

3 ur PCR Reagent s General Applications Complex/ GC-Rich Targets Multiple Targets Routine PCR Reverse Transcription QPCR/ Real Time page 10 Herculase Enhanced DNA s page 11 TaqPlus Maxx DNA page 12 SureStart Taq DNA page 13 StrataScript Reverse Transcriptase page 12 SureStart Taq DNA page 9 PfuTurbo C x Hotstart DNA page 10 Herculase Enhanced DNA s page 12 Taq2000 DNA page 10 Herculase PCR Master Mix page 13 StrataScript RT-PCR Systems page 14 Brilliant QPCR Reagent Kits The Factor Advantage Higher yields Shorter extension times Greater target length capability A key component found exclusively in many of Stratagene s high-fidelity Pfu-based DNA polymerases is the patented polymerase-enhancing factor ***, which improves overall PCR performance. The ArchaeMaxx factor, discovered by Stratagene scientists, eliminates the inhibition of proofreading enzymes caused by incorporation of dutp, which results from deamination of dctp during PCR

4 Stratagene The Leader in High-Performance PCR Expertise Breeds Innovation Our expertise in enzyme engineering keeps us on the leading edge of PCR enzyme development. As the leader in high-fidelity PCR, we offer one of the broadest portfolios of high-fidelity PCR enzymes today. Whether you need the highest accuracy, large product yields, the ability to amplify long or GC-rich targets, or the most reliable enzyme no matter how many targets you have, trust our high-performance polymerases to answer your PCR challenges. High Performance Means High Amplification Efficiency Amplification efficiency is a measure of the percentage of PCR product that doubles with each cycle. Many factors can affect amplification efficiency, including the performance of the polymerase. A slight change in efficiency, magnified over 30 cycles, can have a significant effect on product yield. Quantitative or real-time PCR provides a true determination of amplification efficiency because it can measure results before saturation is reached. Recent testing has shown that our Pfu-based enzymes containing the ArchaeMaxx polymerase-enhancing factor exhibit higher amplification efficiencies than other commercially enzymes 2. Efficiency ND* 0.9 kb 2.6 kb 3.9 kb *ND= Not Detected PfuUltra, PfuTurbo, and Herculase DNA s Pfu DNA Taq DNA Platinum Pfx DNA (Invitrogen) Pfu-Based Enzymes Exhibit Higher Efficiencies Templates of 0.9 kb, 2.6 kb, and 3.9 kb with similar GC-content (53 to 56%) were amplified using various commercially PCR enzymes. Efficiencies shown are averages of at least three independent experiments. Efficiencies of systems using Platinum Pfx DNA polymerase at 2.6 kb and 3.9 kb and Taq DNA polymerase at 3.9 kb could not be detected due to the presence of multiple non-specific products at high concentrations and no measurable product at lower concentrations. 4 P C R R E A G E N T S

5 The Importance of High Fidelity High-fidelity PCR enzymes are valuable for minimizing the introduction of amplification errors in products that will be cloned, sequenced, and expressed. Significant time and effort can be saved by employing high-fidelity amplification procedures that eliminate the need for downstream error-correction steps and minimize the number of clones that must be sequenced in order to obtain errorfree constructs or accurate consensus sequences. Moreover, the use of high-fidelity amplification conditions is essential when analyzing very small amounts of template DNA or rare molecules in heterogeneous populations. Amplifications employing small amounts of template DNA are especially prone to high mutant frequencies, due to PCR-generated errors in early cycles ( jackpot artifacts) and high target doublings 3,4. Is Sequence Verification Breaking Your Bank? Sequence verification is an expensive portion of the total cost of a PCR cloning project. When you use one of our high-fidelity PCR enzymes, you can sequence fewer clones with high confidence that you will obtain your desired product errorfree. The chart below illustrates the probability of an error-free product using various commercially PCR enzymes based on amplicon length and the intrinsic error-rate of each enzyme determined using our published PCR fidelity assay kb amplicon Blunt Mutation % Probability of a Correct Clone if You or 3'-A Frequency Sequence Sequence Sequence PCR Enzyme Ends (%) * 1 Clone 2 Clones 3 Clones PfuUltra Hotstart DNA Blunt PfuTurbo and PfuTurbo C x DNA s Blunt Easy-A High-Fidelity PCR Cloning Enzyme 3'-A Vent DNA Blunt Platinum Pfx DNA Blunt Taq DNA 3'-A kb amplicon Blunt Mutation % Probability of a Correct Clone if You or 3'-A Frequency Sequence Sequence Sequence Sequence Sequence PCR Enzyme Ends (%) * 1 Clone 2 Clones 3 Clones 4 Clones 12 Clones PfuUltra Hotstart DNA Blunt PfuTurbo and PfuTurbo C x DNA s Blunt Easy-A High-Fidelity PCR Cloning Enzyme 3'-A Vent DNA Blunt Platinum Pfx DNA Blunt Taq DNA 3'-A Green shaded areas indicate 99.0% probability of obtaining a correct clone based on the number of clones sequenced. Red shaded areas indicate 99.0% probability of obtaining a correct clone; increased chance that all sequenced clones will contain errors. * Mutation Freq. = (Enzyme Error Rate)*(# bp)*(# doublings) and assumes 106-fold amplification (# doublings = 20) Calculated using the formula: p = 1 (1-f)n where p = probability, f = frequency of obtaining the correct clone, and n = number of clones sequenced w w w. s t r a t a g e n e. c o m 5

6 Highest Accuracy The most accurate PCR enzyme Pfu mutant delivers 300% greater accuracy Ideal for PCR cloning and sitedirected mutagenesis 0-17 kb (genomic) 0-15 kb (vector) Extension time 1 min/kb (0-6 kb) 2 min/kb (>6 kb) Blunt or 3'-A ends Blunt Accuracy vs. Taq 18X factor advantage PfuUltra PfuUltra high-fidelity DNA polymerase *, is an enzyme formulation containing a genetically engineered mutant of Pfu DNA polymerase ** that exhibits enhanced proofreading capability. The accuracy of PfuUltra DNA polymerase was compared to Pfu and Taq DNA polymerases using a validated and referenced fidelity assay 5. Our data demonstrates that PfuUltra high-fidelity DNA polymerase exhibits an average error rate three-fold lower than Pfu DNA polymerase and 18-fold lower than Taq DNA polymerase, making it the most accurate enzyme. The addition of the ArchaeMaxx polymerase-enhancing factor *** promotes higher yields, shorter extension times and greater target length capability. ALSO AVAILABLE: DNA s PfuUltra Hotstart DNA Antibody-based hot-start formulation PfuUltra Hotstart PCR Master Mix 2X formulation including polymerase, buffer, dntps and MgCl 2 M Kb 25 Accuracy 20 Accuracy x PfuUltra High-Fidelity DNA easily amplifies genomic DNA targets up to 17 kb. 5 0 PfuUltra DNA PfuTurbo and PfuTurbo Cx Tgo DNA DNA s Herculase DNA Pfx/KOD DNA s DNA s Expand High Fidelity Advantage-HF Taq DNA Delivers the Greatest Accuracy of Any Available Enzyme PfuUltra high-fidelity DNA polymerase provides the greatest accuracy of any commercially enzyme. Fidelity was measured using Stratagene s validated and referenced fidelity assay. Accuracy = 1/Error Rate. 6 P C R R E A G E N T S

7 Efficient, Accurate TA/UA Cloning Easy-A High-Fidelity PCR Cloning Enzyme Combines the cloning efficiency of Taq DNA polymerase with the accuracy Easy-A high-fidelity PCR cloning enzyme *, is the only proofreading DNA polymerase formulation to offer high-throughput cloning into T- or /U-modified vectors. PCR products amplified with the Easy-A cloning enzyme can be cloned directly, without performing additional steps typically required when amplifying with proofreading polymerases. Adapting blunt-ended fragments amplified with proofreading enzymes requires post-pcr addition of 3'-A overhangs prior to the cloning step. These additional enzymatic steps include a secondary incubation with Taq DNA polymerase and require the opening and closing of tubes, which is detrimental to sample throughput and exposes the lab to contamination risk. Easy-A high-fidelity PCR cloning enzyme does not require any post-pcr A-addition steps. Thus, PCR products generated with the Easy-A PCR cloning enzyme can be added directly to T- or U-modified vectors to provide improved TA/UA cloning without sacrificing accuracy, cloning efficiency, or throughput. ALSO AVAILABLE: Easy-A High-Fidelity PCR Master Mix 2X formulation including polymerase, buffer, dntps and MgCl 2 of Pfu DNA polymerase Proofreading DNA polymerase formulation adds 3'-A overhangs to PCR amplicons as efficiently as Taq High throughput method eliminates post-pcr A-addition steps and contamination risk Hot-start formulation provides enhanced specificity Extension time Blunt or 3'-A ends Accuracy vs. Taq factor advantage 0-5 kb 1 min/kb 3'-A 6X 5' 3' Easy-A -amplified A A PCR products 3' 5' Add µl of PCR product to T-modified vector 5' 3' Easy-A -amplified A A PCR products 3' 5' Add µl of PCR product to U-modified vector T-vector 3' 5' 3' 5' T U T-vector U-vector U-vector T U 5' 3' 5' 3' Topoisomerase-mediated ligation Ligation or conventional ligation T-vector T A PCR product Ligation complete T A T-vector U-vector U A PCR product Ligation complete U A U-vector High Cloning Efficiency with Easy-A PCR Cloning Enzyme Targets amplified with Easy-A PCR Cloning enzyme contain 3'-A overhangs, allowing quick and easy cloning into any T- or U-modified vector. w w w. s t r a t a g e n e. c o m 7

8 High-Fidelity Same high fidelity as Pfu DNA polymerase with superior performance Requires shorter extension times, fewer PCR cycles and less DNA Ideal for amplifying complex genomic DNA targets 0-19 kb (genomic) 0-15 kb (vector) Extension time 1 min/kb (0-6 kb) 2 min/kb (>6 kb) Blunt or 3'-A ends Blunt Accuracy vs. Taq 6X factor advantage PfuTurbo DNA s PfuTurbo DNA polymerase *, is a special formulation of cloned Pfu DNA polymerase ** and the novel polymerase-enhancing factor *** that significantly increases PCR product yields without affecting replication fidelity. The enhanced performance of PfuTurbo DNA polymerase allows the use of shorter extension times, fewer PCR cycles and lower concentrations of DNA template than are required for Pfu DNA polymerase. ALSO AVAILABLE: PfuTurbo Hotstart DNA Antibody-based hot-start formulation PfuTurbo Hotstart PCR Master Mix 2X formulation including polymerase, buffer, dntps and MgCl M PfuTurbo DNA Generates High Yields of Accurate PCR Products PfuTurbo DNA polymerase was used to amplify portions of the human α-1 antitrypsin gene. PCR product was easily generated up to 19 kb. Extension time Blunt or 3'-A ends Accuracy vs. Taq factor advantage 0-4 kb 2 min/kb Blunt 6X Pfu DNA Pfu DNA polymerase *,, derived from the hyperthermophilic archae Pyrococcus furiosus, has been shown to exhibit superior thermostability and proofreading properties compared to other thermostable polymerases. 5 Unlike Taq DNA polymerase, highly thermostable Pfu DNA polymerase possesses 3' to 5' exonuclease proofreading activity that enables the polymerase to correct nucleotidemisincorporation errors. Cloned Pfu DNA polymerase is a recombinant version of native Pfu that provides superior consistency and minimal lot-to-lot variability. 8 P C R R E A G E N T S

9 ...and High Performance PfuTurbo C x Hotstart DNA PfuTurbo Cx hotstart DNA polymerase *, combines high fidelity with trouble-free PCR performance. Formulated with a new mutant of Pfu DNA polymerase, PfuTurbo C x DNA polymerase easily amplifies problematic PCR templates with high accuracy, surpassing other proofreading PCR enzymes in overall performance, and making it the most versatile and reliable proofreading enzyme. Reliable, accurate amplification Tolerates variations in PCR conditions Fidelity equivalent to Pfu and PfuTurbo DNA polymerases Incorporates dutp for use with UNG decontamination protocol Unlike other proofreading PCR enzymes which stop replication just before reaching a uracil residue 6, the novel Pfu mutant in PfuTurbo C x DNA polymerase allows the enzyme to read through a uracil located in the template strand without stalling. Thus PfuTurbo C x DNA polymerase improves the overall reliability of high-fidelity PCR and exhibits less finicky, more robust performance. This product is ideal for amplification of templates of 5 to 10 kb in length, as well as from more difficult systems including targets with high GC content. Although most systems are amplified successfully without the need for additives, DMSO is included with PfuTurbo C x hotstart DNA polymerase to further enhance yields of longer or especially problematic sequences. Extension time Blunt or 3'-A ends Accuracy vs. Taq factor advantage 0-10 kb 1 min/kb (0-6 kb) 2 min/kb (>6 kb) Blunt 6X PfuTurbo Hotstart, 200 µm dttp PfuTurbo C x Hotstart, 200 µm dttp PfuTurbo C x Hotstart, 200 µm dutp PfuTurbo Hotstart, 200 µm dutp PfuTurbo C x Hotstart DNA Successfully Incorporates dutp A 900-bp α-1 anti-trypsin genomic DNA target was amplified with PfuTurbo C x hotstart DNA polymerase and original PfuTurbo hotstart DNA polymerase using nucleotide mixes including 200µM dttp or 200µM dutp. Analysis was performed on Stratagene s Mx3000P realtime PCR system. Data shows no amplification with PfuTurbo DNA polymerase using dutp, but nearly identical amplification results from PfuTurbo C x hotstart DNA polymerase with either dttp or dutp. PfuTurbo Cx AccuPrime Pfx Platinum Pfx 9 kb 6 kb 2.6 kb 0.9 kb Superior Performance with PfuTurbo C x Hotstart DNA Amplification of a range of targets from 0.9 to 9 kb using PfuTurbo C x DNA polymerase and other commercially proofreading PCR enzymes. w w w. s t r a t a g e n e. c o m 9

10 Difficult & GC-Rich Templates / Extra-Long Targets Effectively amplifies problematic templates, including complex and GC-rich Amplifies a broad range of target lengths Higher fidelity than Taq and Taq-based enzyme blends 0-37 kb (genomic) 0-48 kb (vector) Extension time 1 min/kb Blunt or 3'-A ends Mixed Accuracy vs. Taq 3.5X factor advantage Herculase DNA s Herculase enhanced DNA polymerase *, meets today s complex PCR challenges. When you need to accurately amplify a complex or GC-rich template, Herculase DNA polymerase will help you succeed. A unique optimized formulation of high-fidelity Pfu DNA polymerase **, Taq DNA polymerase, and the polymerase-enhancing factor ***, Herculase DNA polymerase excels in amplifying a broad range of target lengths with a more robust yield and superior fidelity than Taq DNA polymerase and other Taq-based blends 7. DMSO is provided separately as a PCR adjunct, and can be added when amplifying difficult targets to increase product yield and extend target-length capability. ALSO AVAILABLE: Herculase Hotstart DNA Antibody-based hot-start formulation Herculase Hotstart PCR Master Mix 2X formulation including polymerase, buffer, dntps and MgCl 2 Competitor s GC-rich PCR Enzyme Herculase +/- DMSO (%) +/- GC-rich solution (M) M Herculase Enhanced DNA Excels in Amplifying Difficult Targets Herculase enhanced DNA polymerase easily amplifies an 82.5% GC-rich fragment of Fragile X gene from human genomic DNA. Successfully amplifies targets >20 kb Higher fidelity than Taq and Taq-based enzyme blends kb (genomic) kb (vector) Extension time 1 min/kb Blunt or 3'-A ends Mixed Accuracy vs. Taq 3.5X factor advantage EXL DNA EXL DNA polymerase *, provides superior performance in amplifying complex targets greater than 20 kb in length, with higher fidelity than Taq DNA polymerase or Taq-based blends. EXL DNA polymerase is a special formulation of Pfu DNA polymerase **, Taq DNA polymerase, and the polymerase enhancing factor ***, optimized specifically for extremely long targets and providing robust yield, specificity, and reliable amplification. EXL Competitor M EXL DNA Excels at Amplifying Extremely Long Targets EXL DNA polymerase easily amplifies two extremely long genomic targets (23 and 30 kb) and a lambda target (45 kb). A competitor s PCR enzyme for long targets was used for comparison. All reactions were performed according to the manufacturer s recommendations. 10 P C R R E A G E N T S

11 Highest PCR Success Rate TaqPlus Maxx DNA TaqPlus Maxx DNA polymerase *, is engineered for maximum PCR reliability. TaqPlus Maxx DNA polymerase is a blend of cloned Taq and Pfu DNA polymerases, and the polymerase-enhancing factor ***. Together with a specially optimized buffer, this enzyme blend provides the highest success rate of any PCR enzyme, including other Taq-based blends. TaqPlus Maxx DNA polymerase reliably produces high PCR product yields on a wide variety of targets up to 10 kb in length. Highest success rate of any PCR polymerase or enzyme blend Consistently high yields on a variety of templates up to 10 kb Greater sensitivity allows successful amplification where starting material is limited High-specificity hot-start allows reliable roomtemperature setup Amplification from small quantities of template can be difficult for most PCR enzymes. TaqPlus Maxx DNA polymerase provides superior PCR sensitivity and can therefore amplify successfully from a fraction of the template quantity required by other polymerases. High sensitivity permits preservation of samples where starting material is limited, and allows TaqPlus Maxx DNA polymerase to detect samples that might otherwise be missed, further adding to its reliability. Extension time Blunt or 3'-A ends Accuracy vs. Taq factor advantage 0-10 kb 1 min/kb Mixed 2X PCR Enzyme Blends Template Qty. (ng) 3.9 kb TaqPlus Maxx Expand High Fidelity Platinum Taq High Fidelity Deoxynucleotide Mix Produces high-quality reaction products Withstands multiple freeze-thaw cycles without compromising efficiency Our dntp mix contains 400 µl of 100 mm dntp mix (25 mm of each dntp), adequate for standard 100 µl primerextension reactions. TaqPlus Maxx DNA Exhibits Superior Sensitivity A 3.9 kb α-1 antitrypsin template was amplified with TaqPlus Maxx DNA polymerase, Expand High Fidelity PCR System and Platinum Taq DNA High Fidelity using decreasing template quantities. 9 kb TaqPlus Maxx PCR Enzyme Blends Expand High-Fidelity Platinum Taq High-Fidelity 4 kb 1.7 kb 0.9 kb TaqPlus Maxx DNA Provides Reliable PCR Results Templates ranging from 0.9 kb to 9 kb were amplified using TaqPlus Maxx DNA polymerase, Expand High Fidelity PCR System and Platinum Taq DNA High Fidelity. All reactions were performed according to the manufacturer s recommendations. w w w. s t r a t a g e n e. c o m 11

12 Routine PCR Hot-start formulation of Taq DNA polymerase Provides greater PCR specificity and yields Reduces nonspecific background Allows reliable roomtemperature setup Extension time Blunt or 3'-A ends Accuracy vs. Taq factor advantage 0-5 kb 1 min/kb 3'-A 1X SureStart Taq DNA SureStart Taq DNA polymerase is a hot-start Taq DNA polymerase. SureStart Taq can be incorporated into PCR protocols previously optimized with Taq DNA polymerase, with little or no modification of cycling parameters or reaction conditions. This specially modified Taq DNA polymerase allows setup of PCR reactions at ambient room temperature without nonspecific annealing and extension during PCR setup, making setup easier. SureStart Taq DNA polymerase can be used in a variety of amplification systems to improve specificity, yield, and amplification of difficult targets. SureStart Taq DNA Increases Specificity A 105 bp fragment of the glucocerebrosidase gene was amplified from human genomic DNA. Reactions were assembled at room temperature and employed 2.5 U of each enzyme and the recommended buffer and cycling parameters. Lane 1: SureStart Taq DNA polymerase, lane 2: unmodified Taq DNA polymerase, lane 3: an antibodybased hot start Taq DNA polymerase, lane 4: competitor s modified Taq DNA polymerase. Ultra-pure cloned Taq DNA polymerase Minimizes smearing Taq2000 DNA Highly thermostable Taq2000 DNA polymerase is a highly purified, recombinant Taq DNA polymerase cloned from the thermophilic eubacteria, Thermus aquaticus. Using Extension time Blunt or 3'-A ends Accuracy vs. Taq factor advantage 0-4 kb 1 min/kb 3'-A 1X Taq2000 DNA polymerase in longer PCR amplifications reduces smearing and virtually eliminates unwanted background artifacts. Taq2000 DNA polymerase has superior thermostability compared to other commercial Taq DNA polymerase preparations. Taq2000 DNA vs. Competitor s Cloned Taq DNA PCR smear artifacts were produced with increasing extension times noted. A 4-kb fragment of transgenic mouse genomic DNA was amplified using Stratagene s Taq2000 DNA polymerase versus another major manufacturer s cloned Taq DNA polymerase. 12 P C R R E A G E N T S

13 RT-PCR StrataScript Reverse Transcriptase StrataScript reverse transcriptase is a Moloney murine leukemia virus reverse transcriptase (MMLV RT) that has been genetically modified to remove RNase H activity. The mutation in the highly conserved residue of the RNase H region results in the loss of RNase H activity without affecting the desired reverse transcriptase function. As a result, StrataScript RT generates longer transcripts and is the preferred enzyme for cdna synthesis. This nuclease-free MMLV RT yields much larger quantities of full-length cdna transcripts than wild-type MMLV RT, which possesses substantial RNase H activity. RNase H deficiency generates longer transcripts Nuclease-free enzyme produces robust cdna yields Ideal enzyme for cdna library construction and RT-PCR 100 U StrataScript RT 100 U SuperScript II RT Same Robust Performance Reactions using 100 U StrataScript reverse transcriptase and 100 U SuperScript II reverse transcriptase generated equivalent performance at 40 C. StrataScript StrataScript first strand cdna synthesis kit employs StrataScript RNase H(-) reverse transcriptase to generate cdna of high quality. Subsequent amplification with sequence-specific primers yields a homogenous population of the specific cdna molecule of interest, eliminating the need for amplification and screening of a library of cdna molecules. StrataScript First Strand cdna Synthesis Kit RT-PCR Systems StrataScript one-tube RT-PCR system includes Easy-A PCR cloning enzyme, a high-fidelity DNA polymerase with terminal transferase activity. Amplified products will contain fewer errors and are ready for subsequent TA/UA cloning with no additional steps required. StrataScript two-tube RT-PCR system includes PfuUltra high-fidelity DNA polymerase, the most accurate PCR enzyme. Both kits contain StrataScript reverse transcriptase. Generates high-quality cdna templates from total or poly(a)+ RNA isolated from a variety of cells and tissue Applies the first-strand synthesis technique for PCR amplification of specific cdna fragments Generates a heterogeneous population of cdna molecules High sensitivity achieved from a wide variety of RNAs Ideally suited for cloning, expression, and sequencing One-tube kit allows quick, sensitive, and reproducible analysis of RNA with convenience and minimal risk of sample contamination Two-tube kit allows archive of cdna samples before amplification step w w w. s t r a t a g e n e. c o m 13

14 QPCR and QRT-PCR Optimized on our Mx4000 and Mx3000P real-time PCR systems Master mixes offer convenience and maximum throughput Core reagent kits allow optimization of reactions SureStart Taq DNA polymerase provides hot-start specificity StrataScript RNase H(-) reverse transcriptase provides superior sensitivity Brilliant QPCR Master Mixes & Core Reagent Kits Brilliant QPCR and QRT-PCR reagents provide maximum versatility in real-time PCR detection. All Brilliant reagent kits contain SureStart Taq DNA polymerase, a hot-start version of Taq that minimizes amplification of non-specific PCR products. Brilliant reagent kits are optimized for use on our Mx4000 and Mx3000P real-time PCR systems, but can be used on virtually any QPCR instrument. Whether you choose to optimize your reactions with a core reagent kit or prefer the convenience of a master mix, there is a Brilliant QPCR or QRT-PCR reagent kit for you. Brilliant probe-based core reagent kits and master mixes are compatible with any fluorescent probe chemistry including TaqMan probes, Molecular Beacons, and Scorpions. Our probe-based core kits are with a standard nucleotide mix or with a GAUC mix ( Plus kits) for dutp/ung decontamination strategy. Brilliant SYBR Green QPCR reagents provide a universal solution to realtime PCR detection and gene quantification, without the need for sequence-specific probes. Choose Brilliant SYBR Green master mixes for maximum convenience, or optimize your reactions using the SYBR Green core reagent kit. Select the Right Brilliant Reagent for You QPCR (DNA) Master Mixes Core Reagent Kits SYBR Green Master Mix Probe-Based Master Mix SYBR Green Core Kit Probe-Based Core Kits (GAUC in Plus kit) QRT-PCR (RNA) Master Mixes Core Reagent Kits SYBR Green Master Mix Probe-Based Master Mix Probe-Based 1-Step Kits (GAUC in Plus kit) Probe-Based 2-Step Kits (GAUC in Plus kit) 14 P C R R E A G E N T S

15 Ordering Information PAGE 6 PfuUltra High-Fidelity DNA 100U U U PfuUltra Hotstart DNA 100U U U PfuUltra Hotstart PCR Master Mix 100 rxn rxn PAGE 7 Easy-A High-Fidelity PCR Cloning Enzyme 100U U U Easy-A High-Fidelity PCR Master Mix 100 rxn rxn PAGE 8 Cloned Pfu DNA 100U U U Native Pfu DNA 100U U U PfuTurbo DNA 100U U U PfuTurbo Hotstart DNA 100U U U PfuTurbo Hotstart PCR Master Mix 100 rxn rxn PAGE 9 PfuTurbo Cx Hotstart DNA 100U U U PAGE 10 EXL DNA 100U U U PAGE 10 continued Herculase Enhanced DNA 100U U U Herculase Hotstart DNA 100U U U Herculase Hotstart PCR Master Mix 100 rxn rxn PAGE 11 TaqPlus Maxx DNA 100U U U PAGE 12 SureStart Taq DNA 100U U U Taq2000 DNA 100U U U PAGE 13 StrataScript First Strand RT-PCR Kit 50 rxn StrataScript One-Tube RT-PCR System 50 rxn StrataScript Reverse Transcriptase 10,000U StrataScript Two-Tube RT-PCR System 50 rxn PAGE 14 Deoxynucleotide Mix 400 µl PROBE-BASED MASTER MIXES Brilliant QPCR Master Mix 200 rxn rxn Brilliant QRT-PCR Master Mix Kit, 1-Step 200 rxn rxn PAGE 14 continued PROBE-BASED CORE REAGENT KITS Brilliant QPCR Core Reagent Kit 200 rxn rxn Brilliant QPCR Plus Core Reagent Kit (dutp/ung) 200 rxn rxn Brilliant QRT-PCR Core Reagent Kit, 1-Step 200 rxn rxn Brilliant QRT-PCR Core Reagent Kit, 2-Step 200 rxn rxn Brilliant QRT-PCR Plus Core Reagent Kit, 1-Step (dutp) 200 rxn rxn Legal Information Scorpions is a trademark of DxS Ltd. PAGE 14 continued Brilliant QRT-PCR Plus Core Reagent Kit, 2-Step (dutp/ung) 200 rxn rxn SYBR GREEN MASTER MIXES Brilliant SYBR Green QPCR Master Mix 200 rxn rxn Brilliant SYBR Green QRT-PCR Master Mix Kit, 1-Step 200 rxn rxn SYBR GREEN CORE REAGENT KITS SYBR is a registered trademark of Molecular Probes. TA Cloning is a registered trademark of Invitrogen Corp. Brilliant SYBR Green QPCR Core Reagent Kit 200 rxn rxn TaqMan is a registered trademark of Roche Molecular Systems, Inc. *U.S. Patent Nos. 6,444,428, 6,379,553, 6,333,165, 6,183,997, 6,489,150, 5,948,663, 5,866,395, 5,556,772, 5,545,552 and patents pending. **U.S. Patent Nos. 6,489,150, 5,948,663, 5,866,395, 5,545,552 and patents pending. ***U.S. Patent Nos. 6,444,428, 6,379,553, 6,333,165, 6,183,997 and patents pending. Purchase of these products is accompanied by a license to use them in the Chain Reaction (PCR) process in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., an authorized thermal cycler. References 1. Hogrefe, et al. (2002). Proc. Nat. Acad. Sci. USA 99: Strategies 16.3, p Hogrefe, H.H. and M.C. Borns. High Fidelity PCR Enzymes. In C.W. Dieffenbach, G.S. Dveksler (eds.) PCR Primer: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., Cha, R.S. and W.G. Thilly. Specificity, efficiency, and fidelity of PCR. In PCR Primer: A Laboratory Manual (eds, Dieffenbach, C.W. and G.S. Dveksler) Cold Spring Harbor Laboratory Press, Cline, J., Braman, J.C. and Hogrefe, H.H. (1996) Nucleic Acids Res. 24: Fogg, et al. (2002). Nat Struct Biol. 9(12): Borns, M. and Hogrefe, H. (2000) Strategies. 13: 12. w w w. s t r a t a g e n e. c o m 15

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