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PMID: 11156625 Published · ppublish English Journal Article

High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers.

Genome research ·Vol. 11 ·No. 1 ·2001-01-00 ·Pages 163-9

Myakishev MV, Khripin Y, Hu S, Hamer DH

Abstract

We have developed a new method for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The technique involves PCR amplification of genomic DNA with two tailed allele-specific primers that introduce priming sites for universal energy-transfer-labeled primers. The output of red and green light is conveniently scored using a fluorescence plate reader. The new method, which was validated on nine model SNPs, is well suited for high-throughput, automated genotyping because it requires only one reaction per SNP, it is performed in a single tube with no post-PCR handling, the same energy-transfer-labeled primers are used for all analyses, and the instrumentation is inexpensive. Possible applications include multiple-candidate gene analysis, genomewide scans, and medical diagnostics.

MeSH Terms
Alleles DNA Primers/chemical synthesis,genetics Energy Transfer/genetics Fluorescein Fluorescent Dyes Genotype Polymerase Chain Reaction/instrumentation,methods Polymorphism, Single Nucleotide/genetics Sensitivity and Specificity Xanthenes
Chemicals
DNA Primers Fluorescent Dyes Xanthenes Texas red Fluorescein
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Myakishev M V
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA. [email protected]
Khripin Y
Hu S
Hamer D H
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Article Info
Journal
Genome research
Abbr.
Genome Res
ISSN
1088-9051
DOI
10.1101/gr.157901
Email
[email protected]
Published
2001-01-00
Pages
163-9
Language
English
Region
United States
NLM ID
9518021
PMCID
PMC311033
Subset
IM
External Links
PubMed source DOI: 10.1101/gr.157901 PMC Full Text View journal details
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