Real-time PCR System

The Real-Time PCR System (also known as Quantitative Real-Time PCR or qPCR) is a cutting-edge molecular biology instrument designed to monitor and quantify DNA amplification during the polymerase chain reaction (PCR) process in real time.

1. Principle and Workflow

Real-Time PCR systems utilize fluorescent dyes or probes (e.g., SYBR Green I, TaqMan probes) to detect DNA amplification as it occurs. Unlike traditional PCR, which measures products at the endpoint, Real-Time PCR tracks fluorescence signals during each cycle, enabling precise quantification of the initial DNA template. The cycle threshold (Ct value), where fluorescence exceeds background levels, correlates directly with the starting DNA concentration.

2. Applications

Gene Expression Analysis: Quantifies mRNA levels to study transcriptional regulation.

Pathogen Detection: Identifies and quantifies viral/bacterial loads (e.g., HIV, HCV, SARS-CoV-2).

Genetic Variation Analysis: Detects SNPs, mutations, and methylation patterns.

Quality Control: Validates gene editing outcomes (e.g., CRISPR) or GMO testing.