11 Facts About RT-PCR

RT-PCR has been used to indicate both real-time PCR and reverse transcription PCR .

Quantification of mRNA using RT-PCR can be achieved as either a one-step or a two-step reaction.

The use of end-point RT-PCR is preferred for measuring gene expression changes in small number of samples, but the real-time RT-PCR has become the gold standard method for validating quantitative results obtained from array analyses or gene expression changes on a global scale.

Measurement approaches of end-point RT-PCR requires the detection of gene expression levels by the use of fluorescent dyes like ethidium bromide, P32 labeling of PCR products using phosphorimager, or by scintillation counting.

End-point RT-PCR is commonly achieved using three different methods: relative, competitive and comparative.

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RT-PCR is widely used in the diagnosis of genetic diseases and, semiquantitatively, in the determination of the abundance of specific different RNA molecules within a cell or tissue as a measure of gene expression.

RT-PCR is commonly used in research methods to measure gene expression.

RT-PCR can be very useful in the insertion of eukaryotic genes into prokaryotes.

RT-PCR can be used to diagnose genetic disease such as Lesch–Nyhan syndrome.

RT-PCR is commonly used in studying the genomes of viruses whose genomes are composed of RNA, such as Influenzavirus A, retroviruses like HIV and SARS-CoV-2.

Quantitative RT-PCR assay is considered to be the gold standard for measuring the number of copies of specific cDNA targets in a sample but it is poorly standardized.