RT Journal
A1 Ranade, Koustubh
A1 Chang, Mau-Song
A1 Ting, Chih-Tai
A1 Pei, Dee
A1 Hsiao, Chin-Fu
A1 Olivier, Michael
A1 Pesich, Robert
A1 Hebert, Joan
A1 Chen, Yii-Der I.
A1 Dzau, Victor J.
A1 Curb, David
A1 Olshen, Richard
A1 Risch, Neil
A1 Cox, David R.
A1 Botstein, David
T1 High-Throughput Genotyping with Single Nucleotide Polymorphisms
JF Genome Research 
JO Genome Research 
YR 2001 
FD July 01 
VO 11 
IS 7 
SP 1262 
OP 1268 
DO 10.1101/gr.157801 
UL http://genome.cshlp.org/content/11/7/1262.abstract 
AB To make large-scale association studies a reality, automated high-throughput methods for genotyping with single-nucleotide polymorphisms (SNPs) are needed. We describe PCR conditions that permit the use of the TaqMan or 5′ nuclease allelic discrimination assay for typing large numbers of individuals with any SNP and computational methods that allow genotypes to be assigned automatically. To demonstrate the utility of these methods, we typed >1600 individuals for a G-to-T transversion that results in a glutamate-to-aspartate substitution at position 298 in the endothelial nitric oxide synthase gene, and a G/C polymorphism (newly identified in our laboratory) in intron 8 of the 11–β hydroxylase gene. The genotyping method is accurate—we estimate an error rate of fewer than 1 in 2000 genotypes, rapid—with five 96-well PCR machines, one fluorescent reader, and no automated pipetting, over one thousand genotypes can be generated by one person in one day, and flexible—a new SNP can be tested for association in less than one week. Indeed, large-scale genotyping has been accomplished for 23 other SNPs in 13 different genes using this method. In addition, we identified three “pseudo-SNPs” (WIAF1161, WIAF2566, and WIAF335) that are probably a result of duplication.