Large-scale genotyping of single nucleotide polymorphisms by Pyrosequencing™ and validation against the 5′nuclease (Taqman®) assay
✍ Scribed by Louise Nordfors; Mårten Jansson; Gunnar Sandberg; Catharina Lavebratt; Selim Sengul; Martin Schalling; Peter Arner
- Publisher
- John Wiley and Sons
- Year
- 2002
- Tongue
- English
- Weight
- 438 KB
- Volume
- 19
- Category
- Article
- ISSN
- 1059-7794
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✦ Synopsis
For the Mutation Detection 2001 Special Issue
Here we present the first large-scale effort at genotyping using a novel sequencing method, Pyrosequencing, as a method for genotyping of single nucleotide polymorphisms (SNPs). Pyrosequencing genotypes were validated through duplicate analysis of 1,022 genotypes using the PSQ96 instrument for pyrosequencing and TaqMan ® for 5¢nuclease assays. Identical results were obtained using both methods. In a small pilot study, a pooling strategy using Pyrosequencing was successfully tested. We conclude that Pyrosequencing is highly efficient and accurate in the analysis of SNPs and represents a promising solution to high-throughput genotyping of large sample populations.