Polymerase chain reaction and its various modifications In: Winter School on Vistas in Marine Biotechnology 5th to 26th October 2010

Thomas, P C (2010) Polymerase chain reaction and its various modifications In: Winter School on Vistas in Marine Biotechnology 5th to 26th October 2010. [Teaching Resource]

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Abstract

Polymerase Chain Reaction or PCR is a molecular technique which allows in vitro synthesis of billions of copies of a target DNA fragment within hours using a simple enzymatic reaction. This is achieved by using a pair oligonucleotide primers that hybridize (anneal) to the opposite strands of the target DNA at positions flanking the region to be amplified. New strands are made through the simultaneous extension of both the primers by addition of nucleotides to the primers. A repetitive series of cycles involving template denaturation, primer annealing and extension of the annealed primers by the enzyme DNA polymerase results in the exponential accumulation of the DNA whose termini are defined by the 5' ends of the primers. Since the primer extension products synthesized in one cycle can serve a template for the next, the number of target DNA copies approximately doubles at every cycle. Thus 20 cycles of PCR can yield about a million-fold amplification. The method is simple, as the PCR can be performed in a single tube. It can be performed on relatively crude DNA containing samples. These factors have made the PCR an attractive method for amplification of specific sequences. This method is extremely rapid; it takes only 3 hours to amplify a known sequence of interest. PCR generates sufficient copy numbers of target DNA sequences for their routine visualization through standard procedures such as electrophoresis followed by staining with ethidium bromide. The PCR products may be sequenced to determine the exact sequence of the nucleotides within the amplified product. As a result, PCR permits routine analysis of DNA from single egg and larvae, and from non-invasively secured tissues such as fin clips and scales. Even partially degraded DNA from poorly preserved sources can be analyzed if sufficiently small PCR products are identified.

Item Type:	Teaching Resource
Uncontrolled Keywords:	Biochemistry; polymerization
Subjects:	Biochemistry Fish Genetics
Divisions:	CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division Subject Area > CMFRI > CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division Subject Area > CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division
Depositing User:	Arun Surendran
Date Deposited:	09 Feb 2023 11:07
Last Modified:	09 Feb 2023 11:07
URI:	http://eprints.cmfri.org.in/id/eprint/16690

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