High-Throughput SNP Genotyping by Allele-Specific PCR with Universal Energy-Transfer-Labeled Primers
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.
Footnotes
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↵3 Corresponding author.
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E-MAIL myakishm{at}mail.nih.gov; FAX (301) 402-3095.
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Article and publication are atwww.genome.org/cgi/doi/10.1101/gr.157901.
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- Received August 2, 2000.
- Accepted October 12, 2000.
- Cold Spring Harbor Laboratory Press
