polymerase chain reaction

The Columbia Encyclopedia, 6th ed.

polymerase chain reaction

polymerase chain reaction (pŏl´Ĭmərās´) (PCR), laboratory process in which a particular DNA segment from a mixture of DNA chains is rapidly replicated, producing a large, readily analyzed sample of a piece of DNA; the process is sometimes called DNA amplification.

The Process

In PCR, DNA (see nucleic acid) is immersed in a solution containing the enzyme DNA polymerase, unattached nucleotide bases (the subunits that DNA is composed of), and "primers," short sequences of nucleotides designed to bind with an end of the desired DNA segment. Two primers are used: one primer binds at one end of the desired segment on one of the two paired DNA strands, and the other primer binds at the other end but on the other strand. The solution is heated to break the bonds between the strands of the DNA. When the solution cools, the primers bind to the separated strands, and DNA polymerase quickly builds a new strand by joining the free nucleotide bases to the primers. When this process is repeated, a strand that was formed with one primer binds to the other primer, resulting in a new strand that is restricted solely to the desired segment. Thus the region of DNA between the primers is selectively replicated. Further repetitions of the process can produce billions of copies of a small piece of DNA in several hours.

Development and Applications

PCR was developed in 1985 by Kary B. Mullis, who was awarded the 1993 Nobel Prize in chemistry for his work. It is used in DNA fingerprinting and in medical tests to identify diseases from the infectious agent's DNA. In forensic use, the test can be used to compare two samples of DNA, usually by looking at matches (or mismatches) of six inherited traits (e.g., hair curliness) from each of the samples. Each trait is controlled by a single gene, each gene having at least two forms, or alleles, resulting in 21 combinations of these alleles, some of them very rare. A nonmatch conclusively excludes a suspect. PCR also is used in taxonomic classification to help show evolutionary relationships between organisms on the molecular level. It has the advantage of being able to be used even when only very small samples, such as tiny pieces of preserved tissue from extinct animals, are available.

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