Reverse transcriptase PCR (RT-PCR) is a laboratory technique used to convert RNA into complementary DNA (cDNA) and then amplify specific DNA targets using polymerase chain reaction (PCR). It is commonly used to measure gene expression and detect RNA viruses.
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RT-PCR involves two main steps: reverse transcription of RNA into cDNA and amplification of the cDNA using PCR.
Reverse transcriptase is the enzyme responsible for synthesizing cDNA from an RNA template.
RT-PCR is crucial in detecting and quantifying viral infections, such as COVID-19, by targeting viral RNA.
The technique can be either one-step or two-step; in one-step RT-PCR, both reverse transcription and PCR occur in a single tube, while in two-step RT-PCR, these processes are performed separately.
Real-time RT-PCR (qRT-PCR) allows for the quantification of RNA levels by measuring the fluorescence emitted during the PCR process.
Review Questions
What are the two main steps involved in RT-PCR?
How does reverse transcriptase function in the RT-PCR process?
Why is RT-PCR particularly useful for detecting RNA viruses?
Related terms
Polymerase Chain Reaction (PCR): A technique used to amplify specific DNA sequences exponentially.
Complementary DNA (cDNA): DNA synthesized from an RNA template by the enzyme reverse transcriptase.
Real-time PCR (qPCR): A variation of PCR that allows for real-time monitoring and quantification of DNA amplification.
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