Open AccessDirect fluorescence analysis of genetic polymorphisms by hybridization with oligonucleotide arrays on glass supports
Author(s) -
Zhen Guo,
Richard A. Guilfoyle,
Andrew J. Thiel,
Renfeng Wang,
Lloyd M. Smith
Publication year - 1994
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/22.24.5456
Subject(s) - oligonucleotide , biology , microbiology and biotechnology , primer (cosmetics) , dna , nucleic acid thermodynamics , polymerase chain reaction , oligomer restriction , biotinylation , exon , primer dimer , gene , genetics , base sequence , multiplex polymerase chain reaction , chemistry , organic chemistry
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.
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