Large-scale genotyping of single nucleotide polymorphisms by Pyrosequencingtrade mark and validation against the 5'nuclease (Taqman((R))) assay

Hum Mutat. 2002 Apr;19(4):395-401. doi: 10.1002/humu.10062.

Abstract

Here we present the first large-scale effort at genotyping using a novel sequencing method, Pyrosequencingtrade mark, as a method for genotyping of single nucleotide polymorphisms (SNPs). Pyrosequencingtrade mark genotypes were validated through duplicate analysis of 1,022 genotypes using the PSQ96trade mark 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 Pyrosequencingtrade mark was successfully tested. We conclude that Pyrosequencingtrade mark is highly efficient and accurate in the analysis of SNPs and represents a promising solution to high-throughput genotyping of large sample populations.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Alleles
  • Apyrase / metabolism
  • Chromosomes, Human, Pair 10 / genetics
  • Genotype
  • Humans
  • Luciferases / metabolism
  • Pilot Projects
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide / genetics*
  • Polymorphism, Single-Stranded Conformational
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods*
  • Sulfate Adenylyltransferase / metabolism
  • Sweden
  • Taq Polymerase / metabolism*
  • Templates, Genetic

Substances

  • Luciferases
  • Taq Polymerase
  • Sulfate Adenylyltransferase
  • Apyrase