Parallel genotyping of over 10,000 SNPs using a one-primer assay on a high-density oligonucleotide array

The analysis of single nucleotide polymorphisms (SNPs) is increasingly utilized to investigate the genetic causes of complex human diseases. Here we present a high-throughout genotyping platform that uses a one-primer assay to genotype over 10,000 SNPs per individual oil a single oligonucleotide array. This approach uses restriction digestion to fractionate the genome, followed by amplification of a specific fractionated Subset of the genome. The resulting reduction in genome complexity enables allele-specific hybridization to the array. The selection of SNPs was primarily determined by computer-predicted lengths of restriction fragments containing the SNPs, and Was further driven by strict empirical measurements Of accuracy, reproducibility, and average call rate, which we estimate to be >9.5%, >99.9%, and >95%, respectively. With average heterozygosity of 0.38 and genome scan resolution of 0.31 cM, the SNP array is a viable alternative to panels of rnicrosatellites (STRs). As a demonstration of the utility of the genotyping platform in whole-genome scans, we have replicated and refined a linkage region on chromosome 2p for chronic mucocutaneous candidiasis and thyroid disease, previously identified using a panel of microsatellite (STR) markers.