- Course Introduction
- 01: Evidence & DNA
- 02: Forensic Biology
- 03: DNA Extraction & Quantitation
- Quantitation
President's DNA Initiative
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Advancing Justice Through DNA Technology
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Advancing Justice Through DNA Technology
DNA Analyst Training
Quantitative PCR
Home > DNA Extraction & Quantitation > Quantitation > Quantitative PCR
Quantitative PCR (qPCR), sometimes refered to as real time PCR, is the most accurate, precise, and efficient method currently available for human DNA quantitation. It is an amplification process that detects and measures the accumulation of fluorescent dyes as the reaction progresses.
The initial quantity of DNA in the sample is detected by monitoring the exponential growth phase of the reaction and measuring the cycle number at which the fluorescent intensity of the sample overcomes the background noise or threshold. This cycle number is directly proportional to the quantity of DNA in the reaction. Analysis of the quantity of DNA in the sample is performed using software that compares the unknowns with the best fit regression line constructed from the standards.
The use of Quantitative PCR is discussed by reference to two methods:
- SYBR® Green detection
- Fluorescent resonance energy transfer (FRET)
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Quantitative PCR instruments include: Applied Biosystems (AB) 7000 and AB 7500, Corbett Rotor-Gene™, Stratagene® Mx 4000®, Cepheid® SmartCycler®, and Roche LightCycler®). 07 |
