Basic Research Workshop

Polymerase chain reaction (PCR)

Kyun-Hwan Kim

Department of Pharmacology, Konkuk University School of Medicine, South Korea

The polymerase chain reaction (PCR) is a biochemical technology to amplify a few copies of DNA generat- ing thousands to millions of copies of a particular DNA sequence in molecular biology. The PCR was invented in 1983 by Kary Mullis and is now a common and often indispensable technique used for a variety of applica- tions in medical and biological research labs. In 1993, Dr. Mullis was awarded the Nobel Prize in Chemistry along with Michael Smith for his work on PCR. Most PCR methods typically amplify DNA fragments of up to ~10 kilo base pairs, although some techni- ques allow for amplification of fragments up to 40 kb in size. The application of PCR include: the selective DNA isolation, DNA cloning for sequencing, functional analysis of genes, amplification and quantification of DNA, diagnosis of hereditary diseases, identification of genetic fingerprints such as forensic sciences and pa- ternity testing, and the detection and diagnosis of infectious diseases (pathogens). Recently, the real-time polymerase chain reaction (RT-PCR), also called quantitative real time polymerase chain reaction (RT-qPCR) or kinetic polymerase chain reaction is widely used to amplify and simultaneously quantify a targeted DNA molecule. Real Time-PCR enables both detection and quantification. The quantifica- tion can be either an absolute number of copies or a relative amount when normalized to DNA input or addi- tional normalizing genes. In this talk, the basic principles, history, and applications of PCR/RT-PCR will be presented.

Keyword: Polymerase chain reaction (PCR), Real-time PCR (RT-PCR), Quantitative RT-PCR (RT-qPCR), Gene amplification.

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