Document Detail


Restriction enzyme analysis of PCR products.
MedLine Citation:
PMID:  19768608     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Progress in single nucleotide polymorphism (SNP) detection technologies has provided information for SNP-based studies, such as identification of candidate genes for the complex genetic diseases, pharmacogenetic analysis, drug development, population genetics, evolutionary studies, and forensic investigations. SNP detection is performed by many methods, including hybridization, allele-specific polymerase chain reaction (PCR), primer extension, oligonucleotide ligation, direct DNA sequencing, and endonuclease cleavage. Each of these methods has its specific advantages and disadvantages. Here we introduce the PCR-restriction fragment length polymorphism (RFLP) method, which has certain advantages over many other techniques used for analysis of SNPs. The PCR-RFLP method allows very rapid, simple, and inexpensive detection of point mutations within the sequences of PCR products. The mutation is discriminated by the specific restriction endonuclease and is identified by gel electrophoresis followed by staining with ethidium bromide. This convenient and simple method is useful in a small basic research study.
Authors:
Masao Ota; Hideki Asamura; Takahito Oki; Masaharu Sada
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  578     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-09-21     Completed Date:  2009-12-30     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  405-14     Citation Subset:  IM    
Affiliation:
Department of Legal Medicine, Shinshu University School of Medicine, Matsumoto, Japan.
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MeSH Terms
Descriptor/Qualifier:
DNA / genetics
DNA Primers / metabolism
DNA Restriction Enzymes / metabolism
HLA-DR Antigens / genetics
Humans
Polymerase Chain Reaction / methods*
Polymorphism, Restriction Fragment Length / genetics
Polymorphism, Single Nucleotide / genetics
Restriction Mapping / methods*
Templates, Genetic
Chemical
Reg. No./Substance:
0/DNA Primers; 0/HLA-DR Antigens; 128338-86-3/HLA-DRB1 antigen; 9007-49-2/DNA; EC 3.1.21.-/DNA Restriction Enzymes

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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