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

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Relation http://eprints.cmfri.org.in/16690/
http://eprints.cmfri.org.in/16686/
 
Title Polymerase chain reaction and its various modifications In: Winter School on Vistas in Marine Biotechnology 5th to 26th October 2010
 
Creator Thomas, P C
 
Subject Biochemistry
Fish Genetics
 
Description 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.
 
Publisher ICAR- Central Marine Fisheries Research Institute
 
Date 2010
 
Type Teaching Resource
NonPeerReviewed
 
Format text
 
Language en
 
Identifier http://eprints.cmfri.org.in/16690/1/Vistas%20in%20Marine%20Biotechnology_2010_Chp%205.pdf
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]