Ver. 2 (EN) All about Real-Time PCR

Transcription

1 Ver. 2 (EN) All about Real-Time PCR 8-11, Munpyeongseoro, Daedeok-gu, Daejeon, 34302, Republic of Korea Tel: (Korea) (International) sales@bioneer.com

2 BIONEER Real-Time PCR Products SYBR Green SYBR Green Real-Time PCR PreMix - GreenStar qpcr PreMix SYBR Green Real-Time PCR MasterMix - 2X GreenStar qpcr Master Mix TaqMan Probe TaqMan Probe Real-Time PCR PreMix - DualStar qpcr PreMix - Plus DualStar qpcr PreMix TaqMan Probe Real-Time PCR Master Mix - Plus DualStar qpcr Master Mix TaqMan Probe Dual HotStart RT-qPCR - Dual-HotStart RT-qPCR PreMix & Master Mix TaqMan Probe - Dual-Labeled Probe Instrument Real-Time PCR Instrument - Exicycler 96 Real-Time Quantitatie Thermal Block Real-Time PCR Reagents Selection Guide Application AccuPower Greenstar qpcr PreMix AccuPower 2X Greenstar Master Mix AccuPower Dualstar qpcr PreMix AccuPower Plus DualStar qpcr AccuPower Dual- HotStart RT-qPCR PreMix Master Mix PreMix Master Mix HotStart PCR SYBR Green I dye Taqman Probe Compatible equipment ABI 7500 Opticon Exicycler 96 ABI 7500 ABI 7500 ABI 7500 All Opticon Opticon All Opticon All Exicycler 96 Exicycler 96 Exicycler 96

3 SYBR Green Method - Real-Time PCR Amplification Reagents Specifications - 5' to 3' exonuclease: No - 3' to 5' exonuclease: No - 3 A Oerhang: Yes - Fragment Size: < 200 bp Application - Real-Time quantification of DNA and cdna targets - Gene expression profiling - Microbial & Viral pathogen detection PreMix Type Cat no. K-6200 K-6201 K-6202 K-6203 K-6204 K-6210 K-6211 K-6212 K-6213 K-6214 Product Description AccuPower GreenStar qpcr PreMix, 50 µl/rxn,12 strips, Exicycler 96, 8-well strip, optical film included AccuPower GreenStar qpcr PreMix, 50 µl/rxn,12 strips, ABI well strip, optical film included AccuPower GreenStar qpcr PreMix, 50 µl/rxn,12 strips, Opticon 8-well strip AccuPower GreenStar qpcr PreMix, 50 µl/rxn, 1 plate, Exicycler 96, 96-well plate, optical film included AccuPower GreenStar qpcr PreMix, 50 µl/rxn,1 plate, ABI well plate, optical film included AccuPower GreenStar qpcr PreMix, 20 µl/rxn,12 strips, Exicycler 96, 8-well strip, optical film included AccuPower GreenStar qpcr PreMix, 20 µl/rxn,12 strips, ABI well strip, optical film included AccuPower GreenStar qpcr PreMix, 20 µl/rxn,12 strips, Opticon 8-well strip AccuPower GreenStar qpcr PreMix, 20 µl/rxn, 1 plate, Exicycler 96, 96-well plate, optical film included AccuPower GreenStar qpcr PreMix, 20 µl/rxn,1 plate, ABI well plate, optical film included Master Mix Type Cat no. K-6251 K-6252 K-6253 K-6254 Product Description AccuPower 2X GreenStar qpcr Master Mix with 50X ROX Dye / 100 rxn, 50 µl reaction AccuPower 2X GreenStar qpcr Master Mix with 50X ROX Dye / 200 rxn, 50 µl reaction AccuPower 2X GreenStar qpcr Master Mix / 100 rxn, 50 µl reaction AccuPower 2X GreenStar qpcr Master Mix / 200 rxn, 50 µl reaction

4 SYBR Green AccuPower GreenStar qpcr PreMix The Real-Time PCR Reagents for a SYBR Green Detection Features and Benefits High Specificity: Non-specific reactions are dramatically eliminated by Enzyme-mediated HotStart Technology (Patent KR , KR ). User Conenience: Just add the template and primers. Extended Stability: Stable for 2 years in a -20 C freezer. Reproducibility: The margin of error is reduced and reproducible results obtained because of the minimal mixing steps. Equipment Compatibility: Generate optimized results on arious real-time PCR instruments such as ABI 7500 fast, Bio-Rad Opticon, and Exicycler 96. Applicable to Various Target Genes: Significant results can be obtained regardless of kinds of target genes. Experimental data The Comparison data with other commercial products Comparison of amplification efficiency between AccuPower GreenStar qpcr PreMix and other company s master mix products. Amplification cure of AccuPower GreenStar qpcr PreMix and other company s Master Mix kit. Lambda DNA primers were added into AccuPower GreenStar qpcr PreMix and other company s Master Mix kit. A series of Lambda DNA positie control diluents were tested. Reaction mixtures were prepared and qpcr was performed according to each company s protocol. All data were obtained using ABI 7500 Fast Real-time PCR machine (Applied Biosystems co.).

5 SYBR Green AccuPower 2X GreenStar qpcr Master Mix Mix Ready-to-use cocktail containing all components, except primer, for the amplification and detection of DNA in real-time quantitatie PCR (qpcr). Features and Benefits High Specificity: AccuPower 2X GreenStar qpcr Master Mix proides more accurate real-time PCR results by the application of HotStart method. Stability: The chemical stabilizer maintains enzyme actiity for 2 years at -20 C. Simplicity: Ready to use, AccuPower 2X Greenstar qpcr Master Mix contains all of the real-time PCR components excluding the primers and template. Reproducibility: Bioneer's strict quality controlled production system ensures that your results are reproducible after experiment Experimental data Using a ABI 7500 Comparison of the specificity of SYBR green assays. Amplification of a 90 bp target gene was detected using serially diluted LP(Legionella Pneumoniae) genomic DNA (10 fold dilution: 10 5 ~10 1 copies) with AccuPower 2X GreenStar qpcr Master Mix and other commercial products. As shown aboe, little amount of primer dimers was appeared in AccuPower 2X GreenStar qpcr Master Mix than other kits.

6 TaqMan probe Method - Real-Time PCR Amplification Reagents Specifications - 5' to 3' exonuclease: Yes - 3' to 5' exonuclease: No - 3 A Oerhang: Yes Application - Gene expression profiling - Target DNA quantification - Microbial detection - Viral/bacterial pathogen load determination AccuPower DualStar qpcr PreMix Cat no. K-6100 K-6101 K-6102 K-6103 K-6104 K-6110 K-6111 K-6112 K-6113 K-6114 Product Description AccuPower DualStar qpcr PreMix, Exicycler 96, 8-well strip, 20 µl, 12 strips, optical film included AccuPower DualStar qpcr PreMix, ABI well strip, 20 µl, 12 strips, optical film included AccuPower DualStar qpcr PreMix, 20 µl, 12 strips, Opticon 8-well strip AccuPower DualStar qpcr PreMix, Exicycler 96, 96-well plate, 20 µl, 1 plate, optical film included AccuPower DualStar qpcr PreMix, ABI well plate, 20 µl, 1 plate, optical film included AccuPower DualStar qpcr PreMix, Exicycler 96, 8-well strip, 50 µl, 12 strips, optical film included AccuPower DualStar qpcr PreMix, ABI well strip, 50 µl, 12 strips, optical film included AccuPower DualStar qpcr PreMix, 50 µl, 12 strips, Opticon 8-well strip AccuPower DualStar qpcr PreMix, Exicycler 96, 96-well plate, 50 µl, 1 plate, optical film included AccuPower DualStar qpcr PreMix, ABI well plate, 50 µl, 1 plate, optical film included AccuPower Plus DualStar qpcr PreMix Cat no. Product Description K-6600 AccuPower Plus DualStar qpcr PreMix, Exicycler 96, 12 strips, Exicycler 8-well strip, 50 µl, optical film included, 96 rxns K-6601 K-6602 AccuPower Plus DualStar qpcr PreMix, ABI 7500, 12 strips, ABI well strip, 50 µl, optical film included, 96 rxns AccuPower Plus DualStar qpcr PreMix,, 50 µl, 12 strips, Opticon 8-well strip, optical film included, 96 rxns AccuPower Plus DualStar qpcr Master Mix Cat no. Product Description K-6603 AccuPower Plus DualStar qpcr Master Mix(2X), 2.5 ml, 100 rxns AccuPower Dual-HotStart RT-qPCR PreMix & Master Mix Cat no. Product Description K-6704 AccuPower Dual-HotStart RT-qPCR PreMix, Exicycler 96, 12 strips, Exicycler 8-well strip, 50 µl, optical film included (96 rxns) K-6705 K-6706 K-6707 AccuPower Dual-HotStart RT-qPCR PreMix, ABI 7500, 12 strips, ABI well strip, 50 µl, optical film included (96 rxns) AccuPower Dual-HotStart RT-qPCR PreMix, Opticon, Opticon 8-well strips, 12 strips, 50 µl, optical film included (96 rxns) AccuPower Dual-HotStart RT-qPCR Master Mix (2X), 2.5 ml, 100 rxns

7 TaqMan probe Method AccuPower DualStar qpcr PreMix The Real-Time PCR Reagents for the Sequence-specific Probe-Based Detection Features and Benefits Specificity: The DualStar qpcr PreMix proides significant reduction of nonspecific reaction, high sensitiity, extended stability and great uniersality. Just additions of primers and fluorescent dye labeled probe of your interesting target gene into DualStar qpcr PreMix proide always reproducible results with conenience of use. User Conenience: Just add probe and primer to your target gene. Extended Stability: Stable for 2 years in a -20 C freezer. Reproducibility: Non-specific reactions are dramatically eliminated by Enzyme-mediated Hotstart Technology (Patent KR , KR ). Equipment Compatibility: Generate optimized results on arious real-time PCR instruments such as ABI 7500 fast, Bio-Rad Opticon, and Exicycler 96. Applicable to Various Target Genes: Significant results can be obtained regardless of kinds of target genes. Experimental data The Comparison data with other commercial products Comparison of detection sensitiity between AccuPower DualStar qpcr PreMix and other supplier s Master Mix products. West Nile Virus (WNV) primers and TaqMan-based probe were added into AccuPower DualStar qpcr PreMix and other supplier s Master Mix kits. A series of WNV positie control diluents were tested. Reaction mixtures were prepared and qpcr was performed according to each supplier s protocol. All data were obtained using ABI 7500 Fast Real-Time PCR system (Applied Biosystems co.).

8 TaqMan probe Method AccuPower Plus DualStar qpcr PreMix & Master Mix Best real-time PCR kits for a dual-probe detection Features and Benefits User Conenience: Just add probe and primers to the PreMix product. With the Master Mix product, you will hae satisfactory results on arious types of qpcr instrument. Dynamic Range: Detect from 10 to 10 8 copies in the same run. Specificity: Obtain the best amplification result with your target gene by using HotStart Taq DNA polymerase. Applicable to Various Target Genes: You can always achiee great real time PCR results een though you use arious types of DNA, cdna, high GC template, and more. Reproducibility: Produced under ISO 9001 Quality Assurance System, AccuPower Plus DualStar qpcr PreMix and 2X Master Mix gie you reproducible results from uniform quality in eery reaction tube. Experimental data 6 Template Con.: 1 x 10 ~ 1 x 10 copies/reaction High Specificity of AccuPower Plus DualStar qpcr PreMix.

9 TaqMan probe Method AccuPower Dual-HotStart RT-qPCR PreMix & Master Mix New technique of one-step RT-qPCR kit Features and Benefits Sensitiity: Obtain results with smallest amount of your target template and high concentrated RNA product. We hae wide dynamic range such as 10 ~ copies. Specificity: This product is idealized to get an accurate target gene by using our premier Dual-HotStart RT-qPCR which is utilizing Pyro-HotStart RT reaction and HotStart PCR. Multiplexing: This product can be used with different dyes (Probe), so you can use arious type of target gene using Exicycler 96 Real-Time Quantitatie Thermal Block from Bioneer. Use of Various Template RNA: You can get satisfying real-time RT-qPCR results since this kit includes Dual-HotStart RTase which proides RT reaction een in high temperature. Therefore, arious template RNA can be used een though it has highly formed secondary structure template RNA. Applicable to Various Types of Samples: This contains arious PCR inhibitors so you can hae accurate RT-qPCR results with template RNA from blood and soil. Easy-to-use: This kit proides AccuPower Dual-HotStart RT-qPCR PreMix or AccuPower Dual-HotStart RT-qPCR Master Mix (2X Master Mix type) which contains thermostable DNA polymerase, RTase, dntps, and more for real-time RT-PCR performance. Just add template RNA, primers, probe, and DEPC distilled water to this kit Reproducibility: Produced under ISO 9001 Quality Assurance System, AccuPower Dual-HotStart RT-qPCR PreMix and 2X Master Mix gie you reproducible results from uniform quality in eery reaction tube. Experimental data High specificity of AccuPower Dual-HotStart RT-qPCR PreMix. Experiment with HCV target and 10 fold serial dilution of template RNA (10 copies ~ 10 6 copies) spiked in Human Total RNA. AccuPower Dual-HotStart RT-qPCR PreMix using Dual-HotStart RT-qPCR method accurately amplifies target RNA without non-specific amplification, een at low concentration of template.

10 TaqMan probe Method TaqMan probe Method - Dual-Labeled Probe for Real-Time PCR Dual-labeled probes, widely-used for real-time qpcr, normally hae a reporter dye at the 5' end and a quencher at the 3' end. Due to its sequence specificity, probes are used for the sensitie quantitatie or qualitatie detection of genes. By attaching different types of fluorophores to the probe, multiplex analysis is also possible to screen multiple genes at once. The major features of Bioneer s dual-labeled probes are as follows: Include free-of-charge serices of Dual-Probe design and Bio-RP purification Proide high ariety of fluorophore-quencher combinations Verify the quality by OD measurement, MALDI-TOF MS, and Fluoroskan test Competitie pricing: Great alue for your research budget Combination of Dual Probe Dye Excitation max (nm) Emission max (nm) Compatible Quencher Dabcyl tamra BHQ 1 BHQ 2 EBQ 6-FAM JOE TET HEX VIC Cy NED TAMRA Cy ROX Texas Red Cy Cy Please inquire Bioneer s technical staff for Dual-Labeled Probe other than those listed aboe.

11 Instrument - Real-Time PCR equipment Exicycler 96 Real-Time Quantitatie Thermal Block Superior 5-color Real-Time Quantitatie PCR system LT Technology Optics to Remoe Well-to-Well Variation To eliminate the well-to-well light intensity ariation associated with internal systems, Bioneer has designed proprietary, patented optics: Light Tunnel technology (Patent KR , US , JP ). Wide Linear Dynamic Range The dynamic range of detection oer 10 9 True 5-Color Multiplexing Due to uneen excitation illumination used in the instruments, separate reference dye must be applied to normalize signal differences between wells by eliminating one of the useful fluorescence channels. Exicycler 96 does not need a reference dye, so all fie channels can be used for 5 color multiplexing. The optics are designed to minimize dye interference, so long-waelength dyes such as Cy5 can be used under the short-arc lamp light source, which emits een illumination throughout the isible spectrum. Filter sets for the most popular dyes are pre-installed, so you don't need to worry about installing more filters. 5-color multiplex PCR data Self-Diagnostics The software performs self instrument diagnostics before each experiment to preent occurring run errors before the experiment. Thus, aluable samples and reagents can be saed from going to waste.

12 Contact Us Bioneer Corporation 8-11 Munpyeongseo-ro, Daedeok-gu Daejeon, 34302, Republic of Korea Tel: (Korea: ) Fax: Bioneer Inc Marina Village PKWY, Suite 110 Alameda, CA 94501, USA Toll Free: Fax: Bioneer R&D Center Korea Bio Park BLDG #B Daewangpangyo-ro, Bundang-gu, Seongnam-si Gyeonggi-do, 13488, Republic of Korea Tel: Fax: