Document Detail


Two-dimensional DNA fingerprinting of human individuals.
MedLine Citation:
PMID:  2565037     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The limiting factor in the presently available techniques for the detection of DNA sequence variation in the human genome is the low resolution of Southern blot analysis. To increase the analytical power of this technique, we applied size fractionation of genomic DNA restriction fragments in conjunction with their sequence-dependent separation in denaturing gradient gels; the two-dimensional separation patterns obtained were subsequently transferred to nylon membranes. Hybridization analysis using minisatellite core sequences as probes resulted in two-dimensional genomic DNA fingerprints with a resolution of up to 625 separated spots per probe per human individual; by conventional Southern blot analysis, only 20-30 bands can be resolved. Using the two-dimensional DNA fingerprinting technique, we demonstrate in a small human pedigree the simultaneous transmission of 37 polymorphic fragments (out of 365 spots) for probe 33.15 and 105 polymorphic fragments (out of 625 spots) for probe 33.6. In addition, a mutation was detected in this pedigree by probe 33.6. We anticipate that this method will be of great use in studies aimed at (i) measuring human mutation frequencies, (ii) associating genetic variation with disease, (iii) analyzing genomic instability in relation to cancer and aging, and (iv) linkage analysis and mapping of disease genes.
Authors:
A G Uitterlinden; P E Slagboom; D L Knook; J Vijg
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  86     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1989 Apr 
Date Detail:
Created Date:  1989-05-24     Completed Date:  1989-05-24     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2742-6     Citation Subset:  IM    
Affiliation:
Department of Molecular Biology, TNO Institute for Experimental Gerontology, Rijswijk, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Blotting, Southern
DNA / genetics*
Electrophoresis, Gel, Two-Dimensional / methods*
Humans
Nucleotide Mapping*
Oligonucleotide Probes
Pedigree
Polymorphism, Genetic*
Polymorphism, Restriction Fragment Length*
Chemical
Reg. No./Substance:
0/Oligonucleotide Probes; 9007-49-2/DNA
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