Genome complexity reduction for SNP genotyping analysis

Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):2942-7. doi: 10.1073/pnas.261710699.

Abstract

Efficient single nucleotide polymorphism (SNP) genotyping methods are necessary to accomplish many current gene discovery goals. A crucial element in large-scale SNP genotyping is the number of individual biochemical reactions that must be performed. An efficient method that can be used to simultaneously amplify a set of genetic loci across a genome with high reliability can provide a valuable tool for large-scale SNP genotyping studies. In this paper we describe and characterize a method that addresses this goal. We have developed a strategy for reducing genome complexity by using degenerate oligonucleotide primer (DOP)-PCR and applied this strategy to SNP genotyping in three complex eukaryotic genomes; human, mouse, and Arabidopsis thaliana. Using a single DOP-PCR primer, SNP loci spread throughout a genome can be amplified and accurately genotyped directly from a DOP-PCR product mixture. DOP-PCRs are extremely reproducible. The DOP-PCR method is transferable to many species of interest. Finally, we describe an in silico approach that can effectively predict the SNP loci amplified in a given DOP-PCR, permitting the design of an efficient set of reactions for large-scale, genome-wide SNP studies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Arabidopsis
  • Base Composition
  • Chromosome Mapping
  • DNA Primers
  • Electrons
  • Gene Library
  • Genome
  • Genotype
  • Humans
  • Mice
  • Physical Chromosome Mapping
  • Polymerase Chain Reaction / methods
  • Polymorphism, Single Nucleotide*

Substances

  • DNA Primers