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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). 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). Combined RT-PCR and qPCR are routinely used for analysis of gene expression and quantification of viral RNA in research and clinical settings.
The use of RT-PCR for the detection of RNA transcript has revolutionized the study of gene expression in the following important ways:
(1)Made it theoretically possible to detect the transcripts of practically any gene
(2Enabled sample amplification and eliminated the need for abundant starting material required when using northern blot analysis
(3)Provided tolerance for RNA degradation as long as the RNA spanning the primer is intact