Reverse transcription polymerase chain reaction

RT-PCR

Reverse transcription polymerase chain reaction (RT-PCR) is a laboratory technique combining reverse transcription of RNA into DNA (in this context called complementary DNA or cDNA) and amplification of specific DNA targets using polymerase chain reaction (PCR).[1] It is primarily used to measure the amount of a specific RNA. This is achieved by monitoring the amplification reaction using fluorescence, a technique called real-time PCR or quantitative PCR (qPCR). Confusion can arise because some authors use the acronym RT-PCR to denote real-time PCR. In this article, RT-PCR will denote Reverse Transcription PCR. Combined RT-PCR and qPCR are routinely used for analysis of gene expression and quantification of viral RNA in research and clinical settings.

The close association between RT-PCR and qPCR has led to metonymic use of the term qPCR to mean RT-PCR. Such use may be confusing,[2] as RT-PCR can be used without qPCR, for example to enable molecular cloning, sequencing or simple detection of RNA. Conversely, qPCR may be used without RT-PCR, for example, to quantify the copy number of a specific piece of DNA.

  1. ^ Freeman WM, Walker SJ, Vrana KE (January 1999). "Quantitative RT-PCR: pitfalls and potential". BioTechniques. 26 (1): 112–22, 124–5. doi:10.2144/99261rv01. PMID 9894600.
  2. ^ Mackay, Ian (2007). Real-time PCR in Microbiology: From Diagnosis to Characterization. Norfolk, England: Caister Academic Press. pp. 440. ISBN 978-1-904455-18-9.