2×SYBR Green qPCR Mix

Storage and Stability

2 x SYBR Green qPCR Mix should be stored protected from light at -20°C for long term storage (more than 6 months), or at 4°C for short term storage. Avoid repeated freezing and thawing.

Description

2 x SYBR Green qPCR Mix provides rapid real-time quantification of RNA targets in SYBR Green fluorescent. The components of 2 x SYBR Green qPCR Mix include Hot-start enzyme named HotMaster Taq DNA Polymerase, dNTP, PCR reaction buffer, BSA and SYBR Green, and so on. The fluorescent dye SYBR Green I in the master mix enables the analysis of many different targets without having to synthesize target specific labeled probes. High specificity and sensitivity in PCR are achieved by the use of the hot-start enzyme HotMaster Taq DNA Polymerase together with a specialized PCR buffer.

Unlike other “hot-start” Taq DNA polymerase formulations that block the enzyme activity only in the first high temperature step, HotMaster Taq DNA inhibitors closes the polymerase-substrate binding site by temperature adjustment method. Inactive polymerase-inhibitor complexes are formed at temperatures below 40°C. As the temperature is elevated to the primer-specific annealing temperature, the binding equilibrium is shifted towards complex formation only with target-specific primed template DNA. This prevents the formation of mis-primed products and primer–dimers during reaction setup and the first denaturation step, leading to high PCR specificity and accurate quantification.

Note

1. Reference Dye ROX is not included in 2 x SYBR Green qPCR Mix. It is for the researcher to decide whether or not ROX reference dye should be included in the experiment, on the basis of technical protocol of qPCR instruments. Its aim is to eliminate the background signal and to adjust and compensate lot-to-lot fluorescence signal error. The catalog number of the supporting ROX product is PC38 ROX reference dye.
2. SYBR® Green I dye is light sensitive and should be stored in the dark, and be protected from light when use.
3. We strongly recommend preparing the PCR reaction on ice, for higher specificity and less background.
4. Mg2+ concentration in 2 x SYBR Green qPCR Mix is 4 mM. We strongly recommend an initial Mg2+ concentration of 2mM. Proper Mg2+ concentration can be achieved by using 25mM MgCl2.