- Quantitative PCR (qPCR) Overview:
- qPCR, also known as Real-time PCR, is a widely used technology for precise analysis of gene expression.
- It can be categorized into dye-based and probe-based methods, with dye-based being more popular, convenient, and cost-effective.
- Dye-based qPCR monitors real-time fluorescence of the dye binding to double-stranded DNA to measure DNA amplification indirectly during each cycle.
- Ct value (Cq value) is determined when the fluorescence signal is distinctly detected over the background, which can be used to evaluate target abundance or calculate absolute quantities.
- SYBR Green 2xqPCR Mix Master Mix:
- Provides superior specificity, robust amplification efficiency, ideal reproducibility, and stability in quantifying target DNA or cDNA.
- It’s a 2X premix containing hot-start Taq polimeraza combined with an antibody for preferable specificity and high amplification speed.
- SYBR Green I in the mix emits fluorescence when bound to double-stranded DNA amplified in each cycle, allowing for indirect quantification of amplification products in real-time.
- Use SYBR or SYBR/FAM scan mode setting on the real-time instrument.
- Limitations of Dye-based qPCR:
- SYBR Green I can intercalate into any double-stranded DNA, including primer-dimers or other undesired products, causing nonspecific fluorescence.
- To confirm specificity, perform a melt curve analysis after amplification, where a sharp peak around the annealing temperature is ideal.
SYBR Green 2xqPCR Mix 1ml *10
$80.00
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