What question did this study set out to answer?

The aim is to develop a real-time quantitative PCR method that improves accuracy and efficiency in measuring gene copies.

October 1, 1996Genome Research6,243 citationsOpen Access

Real time quantitative PCR.

CHChris HeidPromega (United States)JSJunko StevensThermo Fisher Scientific (United States)KLKenneth J. LivakAgave BioSystems (United States)

Key Points

The aim is to develop a real-time quantitative PCR method that improves accuracy and efficiency in measuring gene copies.
Developed a dual-labeled fluorogenic probe method (TaqMan) for quantifying PCR products.
Eliminated post-PCR handling to prevent contamination, increasing throughput.
Demonstrated a dynamic range of target molecule determination spanning five orders of magnitude.
The real-time quantitative PCR method achieved higher accuracy compared to traditional methods.
It showed a significant reduction in labor intensity required for assays.
The new method supports rapid and reproducible gene quantification.

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.

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