In molecular biology, real-time polymerase chain reaction, also called quantitative real time polymerase chain reaction (Q-PCR/qPCR/qrt-PCR) or kinetic polymerase chain reaction (KPCR), is a laboratory technique based on the PCR, which is used to amplify and simultaneously quantify a targeted DNA molecule. For one or more specific sequences in a DNA sample, Real Time-PCR enables both detection and quantification. The quantity can be either an absolute number of copies or a relative amount when normalized to DNA input or additional normalizing genes.
The procedure follows the general principle of polymerase chain reaction; its key feature is that the amplified DNA is detected as the reaction progresses in real time. This is a new approach compared to standard PCR, where the product of the reaction is detected at its end. Two common methods for detection of products in real-time PCR are:
- non-specific fluorescent dyes that intercalate with any double-stranded DNA
- sequence-specific DNA probes consisting of oligonucleotides that are labeled with a fluorescent reporter which permits detection only after hybridization of the probe with its complementary DNA target
Frequently, real-time PCR is combined with reverse transcription to quantify messenger RNA and Non-coding RNA in cells or tissues.
Abbreviations used for real-time PCR methods vary widely and include: RTQ-PCR, Q-PCR or qPCR. Real-time reverse-transcription PCR is often denoted as: qRT-PCR, RRT-PCR, or RT-rt PCR. The acronym "RT-PCR" commonly denotes reverse-transcription PCR and not real-time PCR, but not all authors adhere to this convention.
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