title: High-throughput single nucleotide polymorphism genotyping by fluorescent 5' exonuclease assay.
abstract: Single nucleotide polymorphisms (SNPs) are the class of DNA variants that are most common in the genome. Technologies are needed that allow relatively high-throughput, high-quality genotyping of SNPs, with rapid assay development for individual SNPs of interest. We have implemented an accurate and high-throughput SNP genotyping system using a commercially available fluorescent 5' exonuclease assay. Optimization of the assay system for low-volume reactions and low-probe concentrations has reduced assay costs by 75%. Using a simple assay optimization process, we successfully developed genotyping assays for 92% of assays attempted (309 out of 335) and have generated over 200,000 genotypes.
authors.affiliation: Axys Pharmaceuticals, La Jolla, CA, USA. morin@axyspharm.com
authors.forename: P A
authors.initials: PA
authors.lastname: Morin
authors.forename: R
authors.initials: R
authors.lastname: Saiz
authors.forename: A
authors.initials: A
authors.lastname: Monjazeb
mesh_terms: D000483:Alleles; D004587:Electrophoresis, Agar Gel; D005456:Fluorescent Dyes; D005838:Genotype; D009711:Nucleotides / Q000235:genetics*; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D005456:Fluorescent Dyes; D009711:Nucleotides; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I