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 it 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.
HANBIO's qPCR Premix 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. The qPCR Premix product 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.
| Code NO. | Product Name | Volume |
| 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 |
Simple Operation: The qPCR Premix from Hanbio reduces the number of steps, shorten sample addition time, and minimize contamination risks.
Specificity：Hanbio's qPCR Premix's effectively suppress non-specific amplification and significantly improve amplification efficiency.
Stable and Precise: The qPCR Premix is capable of accurate quantification across a wide range of template concentrations, yielding stable and reliable qPCR results.
| 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.
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.
The key components typically include a DNA polymerase, dNTPs, MgCl2, a buffer system, and sometimes a fluorescent dye for detection.
Hanbio's qPCR Premix is a ready-to-use solution designed for quantitative PCR (qPCR) applications. It contains all the necessary components for qPCR, including DNA polymerase, dNTPs, MgCl2, and optimized buffer, except for the template DNA and primers.
The main applications include gene expression analysis, quantification of DNA or RNA, genotyping, and detection of pathogens.
The qPCR Premix should be stored at -20°C. Avoid repeated freeze-thaw cycles to maintain its performance.
The recommended reaction volume is usually 20 µL or 25 µL, but it can vary depending on the specific protocol and equipment used.
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