Open AccessHigh-Throughput Single Nucleotide Polymorphism Genotyping by Fluorescent 5′ Exonuclease Assay
Author(s) -
Phillip A. Morin,
Robert Saiz,
Atousa Monjazeb
Publication year - 1999
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/99273rr02
Subject(s) - genotyping , snp genotyping , single nucleotide polymorphism , molecular inversion probe , exonuclease , biology , genotype , microbiology and biotechnology , snp , genetics , dna , gene , dna polymerase
Single nucleotide polymorphisms (SNPs) are the class of DNA variants that are most common in the genome. Technologies are needed that allow relatively highthroughput, 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.