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Published online before print April 14, 2003, 10.1101/gr.939903
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Vol 13, Issue 5, 925-931, May 2003

METHODS

Exo-Proofreading, A Versatile SNP Scoring Technology

Patrick Cahill1, Michele Bakis, James Hurley, Veena Kamath, William Nielsen, Dina Weymouth, Josée Dupuis, Lynn Doucette-Stamm and Douglas R. Smith

Genome Therapeutics Corporation, Waltham, Massachusetts 02453, USA

We report the validation of a new assay for typing single nucleotide polymorphisms (SNPs) that takes advantage of the 3'-to-5' exonuclease proofreading activity of many DNA polymerases. The assay uses one or more primers labeled on the 3' nucleotide base, and can be implemented in a variety of formats including a one-step PCR reaction that allows SNP typing directly from genomic DNA samples. The detection of genotypes can be accomplished by means of fluorescence detection on assays that have been purified to remove excess primer, or by means of fluorescence polarization without any additional cleanup. We also demonstrate that the Exo-Proofreading SNP assay can be used on pooled samples to obtain allele frequency data.

1 Corresponding author.

E-MAIL pcahill{at}genomecorp.com; FAX (781) 398-2472.

Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.939903. Article published online before print in April 2003.


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D. A. Di Giusto and G. C. King
Strong positional preference in the interaction of LNA oligonucleotides with DNA polymerase and proofreading exonuclease activities: implications for genotyping assays
Nucleic Acids Res., February 18, 2004; 32(3): e32 - e32.
[Abstract] [Full Text] [PDF]


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