Large-scale genotyping of single nucleotide polymorphisms by Pyrosequencing (TM) and validation against the 5 ' nuclease (TaqMan (R)) assay

Here we present the first large-scale effort at genotyping using a novel sequencing method, Pyrosequencing(TM), as a method for genotyping of single nucleotide polymorphisms (SNPs). Pyrosequencing(TM) genotypes were validated through duplicate analysis of 1,022 genotypes using the PSQ96(TM) instrument for pyrosequencing and TaqMan(R) for 5'nuclease assays. Identical results were obtained using both methods. In a small pilot study, a pooling strategy using Pyrosequencing(TM) was successfully tested. We conclude that Pyrosequencing(TM) is highly efficient and accurate in the analysis of SNPs and represents a promising solution to high,throughput genotyping of large sample populations. Hum Mutat 19:395-401, 2002. (C) 2002 Wiley-Liss, Inc.