title: Exo-proofreading, a versatile SNP scoring technology.
abstract: 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.
authors.affiliation: Genome Therapeutics Corporation, Waltham, Massachusetts 02453, USA. pcahill@genomecorp.com
authors.forename: Patrick
authors.initials: P
authors.lastname: Cahill
authors.forename: Michele
authors.initials: M
authors.lastname: Bakis
authors.forename: James
authors.initials: J
authors.lastname: Hurley
authors.forename: Veena
authors.initials: V
authors.lastname: Kamath
authors.forename: William
authors.initials: W
authors.lastname: Nielsen
authors.forename: Dina
authors.initials: D
authors.lastname: Weymouth
authors.forename: Josee
authors.initials: J
authors.lastname: Dupuis
authors.forename: Lynn
authors.initials: L
authors.lastname: Doucette-Stamm
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Smith
mesh_terms: D000483:Alleles; D004252:DNA Mutational Analysis / Q000379:methods* / Q000592:standards*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D005787:Gene Frequency / Q000235:genetics; D006801:Humans; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods / Q000592:standards
chemical_list: C118225:Pwo polymerase; D004259:DNA-Directed DNA Polymerase