Real time quantitative PCR.

Chris Heid, Junko Stevens, Kenneth J. Livak, P. Williams

Genome Research · 1996 · 6.2K citations · 33 references

DOIFull text

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Concepts

TL;DR

It uses a dual‑labeled fluorogenic probe (TaqMan) to monitor PCR product accumulation in real time. The real‑time PCR method accurately and reproducibly quantifies gene copies over a five‑order‑of‑magnitude dynamic range, eliminates post‑PCR handling to reduce contamination, and offers higher throughput and lower labor than existing quantitative PCR techniques.

Abstract

We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe (i.e., TaqMan Probe). This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over contamination and resulting in much faster and higher throughput assays. The real-time PCR method has a very large dynamic range of starting target molecule determination (at least five orders of magnitude). Real-time quantitative PCR is extremely accurate and less labor-intensive than current quantitative PCR methods.

References

33