Discussion board for any problem in quantitative real-time PCR.
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- the reference in real time PCR, the academic and industrial information platform. The GQ pages describe and summarize all technical aspects involved in quantitative gene expression analysis using real-time qPCR, digital PCR & RT-qPCR.
In Press, Corrected Proof, Available online 29 April 2017
Amin Forootan, Robert Sjöback, Jens Björkman, Björn Sjögreen, Lucas Linz, Mikael Kubista
Abstract: Quantitative Real-Time Polymerase Chain Reaction, better known as qPCR, is the most sensitive and specific technique we have for the detection of nucleic acids. Even though it has been around for more than 30 years and is preferred in research applications, it has yet to win broad acceptance in routine practice. This requires a means to unambiguously assess the performance of specific qPCR analyses. Here we present methods to determine the limit of detection (LoD) and the limit of quantification (LoQ) as applicable to qPCR. These are based on standard statistical methods as recommended by regulatory bodies adapted to qPCR and complemented with a novel approach to estimate the precision of LoD. Keywords: Limit of detection; Limit of quantification; LoD; LoQ; GenEx software; Real-time PCR; qPCR; Data analysis; Replicates; MIQE; Quality control; Standardization
Read the BDQ Special Issue on digital PCR. The articles in this Special Issue illustrate the potential of this powerful technique to advance clinical research and routine diagnosis with its reproducibility and improved sensitivity.