汉恒生物-qPCR Premix

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qPCR Premix

HBStart Green qPCR Premix is a ready-to-use master mix supplied at 2× concentration, consisting of DNA polumerase engineered with a proprietary hot-start mechanism, SYBR Green I dye, dNTP, buffer components, etc. The user only needs to template, provide primers, and water.

The chemically modified DNA polymerase is equipped with a proprietary hot-start mechanism that completely inhibits enzyme activity at room temperature to provides improved specificity. The polymerase is re-activated after a few minutes incubation at 95°C.

Free or single-stranded DNA-bound SYBR Green I has a low level of intrinsic fluorescence; however, SYBR Green I exhibits optimal fluorescence when bound to double-stranded DNA. So i t can be used on the quantitative detection of target gene based on the principle that an increase in fluorescence intensity is proportional to the amount of double-stranded PCR product produced.

Product packaging and storage conditions ：

This product is stored at -20℃ away from light and used within one year. For frequent use in the short term, the premix can be stored at 4℃ away from light. It has been tested that storing the product at 4℃ for a week, or repeatedly freezing and thawing 10 times, does not affect the use of the effect.

Product Number	Product Name	Pack Size
HB-START-1000	HBStart Green qPCR Premix	1mL
HB-START(LR)-1000	HBStart Green qPCR Premix (Low Rox)	1mL
HB-START(HR)-1000	HBStart Green qPCR Premix (High Rox)	1mL

Method：

1、PCR Reaction Mixture

Component	Volume(10ul)	Volume(20ul)	Final Concentration
Template	XuL	XuL
Forward Primer (10 uM)	0.2uL	0.4uL	0.2uM
Reverse Primer (10 uM)	0.2uL	0.4uL	0.2uM
2×HBStart Green qPCR Premix	5uL	10uL	1X
Nuclease-free Water	（4.6-X）uL	（9.2-X）uL
Total Volume	10uL	20uL

Note：

I. It is recommended to apply DNA template in quantities less than 100 ng. Since the copy number of different types of template genes varies, the optimal amount could be determined by developing a gradient dilution series of tests.

II. DNA template should be added in less than 10% the volume of the PCR reaction mixture.

2、PCR protocol

Two-step RT-qPCR is best suited for applications requiring high specificity, while three-step

TR-qPCR is suited for applications requiring high amplification efficiency.