Document Detail


Polymerase chain reaction : basic protocols.
MedLine Citation:
PMID:  21400259     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
The melding of a technique for repeated rounds of DNA synthesis with the discovery of a thermostable DNA polymerase has given scientists the very powerful technique known as polymerase chain reaction (PCR). PCR is based on three simple steps required for any DNA synthesis reaction: (1) denaturation of the template into single strands; (2) annealing of primers to each original strand for new strand synthesis; and (3) extension of the new DNA strands from the primers. These reactions may be carried out with any DNA polymerase and result in the synthesis of defined portions of the original DNA sequence. However, in order to achieve more than one round of synthesis, the templates must again be denatured, which requires temperatures well above those that inactivate most enzymes. Therefore, initial attempts at cyclic DNA synthesis were carried out by adding fresh polymerase after each denaturation step (1,2). The cost of such a protocol becomes rapidly prohibitive.
Authors:
B C Delidow; J P Lynch; J J Peluso; B A White
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  15     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  1993  
Date Detail:
Created Date:  2011-03-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1-29     Citation Subset:  IM    
Affiliation:
Department of Anatomy, University of Connecticut Health Center, Farmington, CT.
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