Genotyping single nucleotide polymorphisms directly from genomic DNA by invasive cleavage reaction on microspheres.

Abstract:

:Here we report proof-of-principle for a microsphere-based genotyping assay that detects single nucleotide polymorphisms (SNPs) directly from human genomic DNA samples. This assay is based on a structure-specific cleavage reaction that achieves single base discrimination with a 5'-nuclease which recognizes a tripartite substrate formed upon hybridization of target DNA with probe and upstream oligonucleotides. The assay is simple with two easy steps: a cleavage reaction, which generates fluorescent signal on microsphere surfaces, followed by flow cytometry analysis of the microspheres. Genomic DNA samples were genotyped for the SNP in the Apolipoprotein E gene at amino acid position 158. The assay successfully scored wild type, heterozygous and homozygous mutants. To our knowledge, this is the first report of a solid-support assay for detection of SNPs directly from genomic DNA without PCR amplification of the target.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Rao KV,Stevens PW,Hall JG,Lyamichev V,Neri BP,Kelso DM

doi

10.1093/nar/gng066

keywords:

subject

Has Abstract

pub_date

2003-06-01 00:00:00

pages

e66

issue

11

eissn

0305-1048

issn

1362-4962

journal_volume

31

pub_type

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