Tools for genetics and genomics: Polymerase chain reaction
- Benjamin A Raby, MD, MPH
Benjamin A Raby, MD, MPH
- Section Editor — Genetics
- Associate Professor of Medicine
- Harvard Medical School
The polymerase chain reaction (PCR) is the basis of many modern molecular biology and molecular genetics techniques. In just a few hours, PCR can amplify a single DNA molecule a million-fold . The greatly amplified target DNA is subsequently analyzed via other techniques.
Since its first publication in 1985 , the impact of PCR on biomedical research has been immense. This technology allows large quantities of rare sequences to be synthesized, cloned, and analyzed with high reliability and minimum effort. The award of the 1993 Nobel Prize in Chemistry to Kary B. Mullis for inventing the technique recognized the importance of PCR-based methods .
PCR allows for rapid and highly-specific amplification of DNA fragments. The method is relatively inexpensive and commonly performed in most molecular laboratories. Pieces of DNA from about 50 base pairs (bp) to over 10 kilobases (kb) can be amplified, even from vanishingly small amounts of starting genomic DNA. The two most important principles underlying PCR are:
●Complementarity-driven binding of single DNA strands to form a duplex
●Template-driven, semi-conservative synthesis of DNA, by DNA polymerases
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- THE PCR PROCESS
- UNIQUE GENOMIC SEQUENCES
- PCR COMPARED TO OTHER AMPLIFICATION METHODS
- Whole genome amplification
- RNA amplification
- CLINICAL APPLICATIONS OF PCR
- Genotyping and sequencing
- - SNP genotyping
- - Detection of rare sequences
- Quantifying the amount of a nucleic acid sequence
- - RT-PCR
- - Digital PCR (dPCR)
- - Gene expression profiles
- - Clinical measurement of viral RNA or DNA
- CRITICAL EVALUATION OF DATA