The system contains sufficient reagents suitable for 200 amplification reactions of 50 μl PCR system.
This reagent can be stored at 4°C for 2 months and protected from light. For longer storage, this reagent should be kept at -20°C and protected from light.
2x SYBR® Green qPCR Mix (High Rox+) is designed for high-performance, high-throughput, real-time PCR. The kit contains a Taq DNA polymerase engineered through a process of molecular evolution. The result is a unique enzyme, specifically designed for qPCR using SYBR® Green I dye chemistry.
2x SYBR® Green qPCR Mix (High Rox+) is a convenient premix of the components (except primers, template, and water) necessary to perform real-time polymerase chain reaction (PCR) using SYBR® Green I dye with enhanced sensitivity and specificity. The SYBR® Green dye binds to double-stranded (ds) DNA, thus providing a fluorescent signal that reflects the amount of dsDNA product generated during PCR. This reagent is used in amplification and detection of DNA in qPCR on ABI real-time instruments that support normalization with high Rox reference dye at a final concentration of 500 nM.
• This reagent can be used in ABI Real-time systems that require high concentration of Rox reference dye.
• Hot Start technology with anti-Taq DNA polymerase antibodies enables high specificity and reproducible amplification.
Composition of the SYBR Green qPCR Mix
100 mM KCl , 5 mM MgCl2, 400 μM dNTPs, 0.1 U/μl Hot Start Taq DNA Polymerase, 1x SYBR® Green, 2% Rox reference dye and other optimized buffer components.
• This reagent can be used in general detection devices, such as: LineGene (Bioer Technology co., ltd.)
• This reagent can also be used in detection equipment using glass capillaries or passive reference, such as:
LightCycler (Roche Molecular Systems)
ABI PRISM® 7000, 7700, and 7900 (Applied Biosystems)
Note: The passive reference mode of detectors should be set at“ROX”.
Guidelines for preventing contamination of PCR reaction
During PCR more than 10 million copies of template DNA are generated. Therefore, care must be taken to avoid contamination with other templates and amplicons that may be present in the laboratory environment. General recommendations to lower the risk of contamination are as follows:
• Prepare your DNA sample, set up the PCR mixture, perform thermal cycling and analyze PCR products in separate areas.
• Set up PCR mixtures in a laminar flow cabinet equipped with an UV lamp.
• Wear fresh gloves for DNA purification and reaction set up.
• Use reagent containers dedicated for PCR. Use positive displacement pipettes, or use pipette tips with aerosol filters to prepare DNA samples and perform PCR set up.
• Always perform “no template control” (NTC) reactions to check for contamination
The absence of endodeoxyribonucleases, exodeoxyribonucl- eases and ribonucleases is confirmed by appropriate quality tests. Functionally tested in amplification of a single-copy gene from human genomic DNA.