Document Detail


Optimization of nonradioisotopic single strand conformation polymorphism analysis with a conventional minislab gel electrophoresis apparatus.
MedLine Citation:
PMID:  8238876     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have developed a means of nonradioisotopic single strand conformation polymorphism (nonRI-SSCP) analysis and applied it to the detection of a point mutation in the human tumor suppressor gene, p53. The method does not require any particular facilities or apparatus, such as a laboratory for radioactive materials, a large gel unit for sequencing, or a semiautomated electrophoresis system. This technique comprises amplification of DNA fragments by the PCR technique with specific oligonucleotide primers, denaturation, and electrophoresis on neutral polyacrylamide gels in a conventional minislab apparatus. The SSCP patterns on electrophoresis were detected with a commercially available silver stain method. We also evaluated various electrophoretic conditions for nonRI-SSCP analysis, such as the gel concentration and buffer components. A tris/glycine buffer system gave better resolution of SSCP bands. The SSCP patterns of different sized DNAs could be analyzed in a gradient polyacrylamide gel. Thus, nonRI-SSCP analysis with a conventional minislab gel electrophoresis apparatus can be satisfactorily substituted for a commonly used RI-SSCP technique.
Authors:
M Oto; S Miyake; Y Yuasa
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Analytical biochemistry     Volume:  213     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1993 Aug 
Date Detail:
Created Date:  1993-12-03     Completed Date:  1993-12-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  19-22     Citation Subset:  IM    
Affiliation:
Department of Biotechnology, Tokyo Technical College, Japan.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
DNA, Neoplasm / analysis*,  genetics
DNA, Single-Stranded / analysis*
Electrophoresis, Polyacrylamide Gel / methods
Genes, p53 / genetics
Humans
Molecular Sequence Data
Nucleic Acid Conformation
Polymerase Chain Reaction / methods
Polymorphism, Genetic*
Sensitivity and Specificity
Silver Staining / methods
Templates, Genetic
Chemical
Reg. No./Substance:
0/DNA, Neoplasm; 0/DNA, Single-Stranded

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


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