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Application: Useful for quantification of DNA.
Background: Quantitative PCR (qPCR), also known as real-time PCR, monitors the amplification of a DNA fragment throughout the reaction using a fluorescent dye. It is a tool with high sensitivity, fast turnaround time, and minimal risk of contamination. Commonly used dyes include Thiazole green, which binds to the minor groove of double-stranded DNA (dsDNA) and fluoresces over 1000-fold. This fluorescence allows to quantify the amount of product during any given cycle. The baseline threshold, which is usually set above background fluorescent noise levels, is used to calculate the cycle threshold (Ct or Cq value) and quantitatively evaluate the results. The Ct/Cq values can be used to determine the starting amounts of DNA using a standard curve, and the melting curves can help assess quality of the amplified product. qPCR is a standard technique in gene expression analysis, mRNA analysis, copy number variation (CNV) analysis, and pathogen detection, among others. Its value in medicine and biology makes it crucial in any research workflow.
Description: The WaveQuantTM 2x qPCR Master Mix is designed to provide a high-quality and reliable real-time qPCR (quantitative polymerase chain reaction)-based platform for sensitive detection of gene expression. It comes conveniently pre-mixed, allowing for a quick experimental setup, and optimized for use with double-stranded DNA. This mix contains Taq polymerase, Mg2+, and the fluorescent dye Thiazole green, which is structurally identical to SYBR® Green I. The dye can bind multiple times to a single DNA strand to enhance sensitivity.This product has been validated using two housekeeping genes as controls.Figure 1. Principle of dye-based qPCR. Thiazole Green binds to newly formed dsDNA during the qPCR cycles, allowing for real-time monitoring and quantification.
Storage Stability: This mix will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: WaveQuantTM 2x qPCR Master Mix supplied as 2.5 ml, sufficient for 250 reactions.
Unit Definition: Reagent
Biosafety Level: Not applicable (BSL-1)
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