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What is the difference between RT-PCR, QPCR, and Real-time PCR?


What is the difference between RT-PCR, QPCR, and Real-time PCR?

What is RT-PCR?

RT-PCR is reverse transcription PCR (reverse transcription PCR) or reverse transcription-PCR (RT-PCR), which is a widely used variant of polymerase chain reaction (PCR). In RT-PCR, an RNA strand is reverse transcribed into complementary DNA, which is then used as a template for DNA amplification by PCR.

What is qPCR, Real-time-PCR?

In fact, Realtime-PCR and qPCR (Quantitative Rea-ltime-PCR) are the same thing, both are real-time quantitative PCR, which means that each cycle in the PCR process has a real-time record of data, so the number of starting templates can be analyzed. Precise analysis.

Although Real-time PCR (real-time fluorescent quantitative PCR) and Reverse transcription PCR (reverse transcription PCR) seem to be abbreviated as RT-PCR. However, the international convention is: RT-PCR refers specifically to reverse transcription PCR, Real-time PCR is generally abbreviated as qPCR (quantitative real-time PCR).

What is real-time RT-PCR (or RT-qPCR)?

The "RT" in the middle of real-time RT-PCR is reverse transcription (reverse transcription), and the whole meaning is the real-time reverse transcription PCR technology (quantitative Real-time RT-PCR) using mRNA or total RNA as a template. Real time RT-PCR refers to the combination of qPCR + RT-PCR, which means that mRNA is reverse transcribed into cDNA and then used as a template for real-time fluorescent PCR analysis, because RT-PCR can be qualitative, but not quantitative detection. .

Therefore, it is not difficult to understand that real-time RT-PCR (RT-qPCR) is a reverse transcription PCR combined with fluorescence quantitative technology: first, cDNA (RT) is obtained from RNA reverse transcription, and then quantitative analysis is performed by Real-time PCR ( qPCR).

Therefore, real-time fluorescence quantitative PCR, it can not only use cDNA as a template, but also use genomic DNA as a template.


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