TaqMan genotyping of insertion/deletion polymorphisms

Methods Mol Biol. 2005:311:165-76. doi: 10.1385/1-59259-957-5:165.

Abstract

The 5' fluorogenic (TaqMan) assay has been successfully used in screening for single-nucleotide polymorphisms; the very few steps required and the ability to automate each step allow for high-throughput screening. Insertion/deletion polymorphisms are an important class of markers that can be studied for different applications, such as diagnostics, genome variation, and species identification. Polymerase chain reaction (PCR) and post-PCR analysis are required to score the insertion or the deletion allele. In this chapter, we describe an expansion of the TaqMan technology for a rapid, high-throughput, screening for insertion/deletion polymorphisms in which the exact endpoints are known. The method requires minimal post-PCR analysis and can be applied to polymorphisms of any size.

Publication types

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

MeSH terms

  • Alleles
  • DNA Primers
  • DNA Probes
  • Genotype
  • Humans
  • Mutagenesis, Insertional*
  • Polymorphism, Single Nucleotide*
  • Sequence Deletion*
  • Taq Polymerase / genetics*

Substances

  • DNA Primers
  • DNA Probes
  • Taq Polymerase