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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 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 |
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.
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