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PMID: 7816638 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Direct fluorescence analysis of genetic polymorphisms by hybridization with oligonucleotide arrays on glass supports.

Nucleic acids research ·Vol. 22 ·No. 24 ·1994-12-11 ·Pages 5456-65

Guo Z, Guilfoyle RA, Thiel AJ, Wang R, Smith LM

Abstract

A simple and rapid method for the analysis of genetic polymorphisms has been developed using allele-specific oligonucleotide arrays bound to glass supports. Allele-specific oligonucleotides are covalently immobilized on glass slides in arrays of 3 mm spots. Genomic DNA is amplified by PCR using one fluorescently tagged primer oligonucleotide and one biotinylated primer oligonucleotide. The two complementary DNA strands are separated, the fluorescently tagged strand is hybridized to the support-bound oligonucleotide array, and the hybridization pattern is detected by fluorescence scanning. Multiple polymorphisms present in the PCR product may be detected in parallel. The effect of spacer length, surface density and hybridization conditions were evaluated, as was the relative efficacy of hybridization with single or double-stranded PCR products. The utility of the method was demonstrated in the parallel analysis of 5 point mutations from exon 4 of the human tyrosinase gene.

MeSH Terms
Alleles Amino Acid Sequence Base Sequence Cross-Linking Reagents Exons/genetics Fluorescein Fluoresceins Glass Humans Image Processing, Computer-Assisted Molecular Probe Techniques Molecular Sequence Data Monophenol Monooxygenase/genetics Nucleic Acid Hybridization Oligonucleotide Probes/chemical synthesis,isolation & purification Point Mutation Polymerase Chain Reaction Polymorphism, Genetic
Chemicals
Cross-Linking Reagents Fluoresceins Oligonucleotide Probes Monophenol Monooxygenase Fluorescein
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Guo Z
Department of Chemistry, University of Wisconsin, Madison 53706-1396.
Guilfoyle R A
Thiel A J
Wang R
Smith L M
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/22.24.5456
Published
1994-12-11
Pages
5456-65
Language
English
Region
England
NLM ID
0411011
PMCID
PMC332096
Subset
IM
Grants
NHGRI NIH HHS · 2R01HG00321 · United States
NIGMS NIH HHS · GM08293-05 · United States
Databases
GENBANK
M63238
External Links
PubMed source DOI: 10.1093/nar/22.24.5456 PMC Full Text View journal details
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