Unit

UNIT 2.10 Genotyping Using the TaqMan Assay

  1. Lester Hui,
  2. Terrye DelMonte,
  3. Koustubh Ranade

Published Online: 1 JAN 2008

DOI: 10.1002/0471142905.hg0210s56

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Hui, L., DelMonte, T. and Ranade, K. 2008. Genotyping Using the TaqMan Assay. Current Protocols in Human Genetics. 56:2.10:2.10.1–2.10.8.

Author Information

  1. Bristol-Myers Squibb, Princeton, New Jersey

Publication History

  1. Published Online: 1 JAN 2008
  2. Published Print: JAN 2008

Abstract

The 5′-nuclease allelic discrimination assay, or TaqMan assay, is a PCR-based assay for genotyping single nucleotide polymorphisms (SNPs). The region flanking the SNP is amplified in the presence of two allele-specific fluorescent probes. The probes do not fluoresce in solution because of a quencher at the 3′ end. The presence of two probes allows the detection of both alleles in a single tube. Moreover, because probes are included in the PCR, genotypes are determined without any post-PCR processing, a feature that is unavailable with most other genotyping methods. This unit describes probe and primer design and PCR conditions. Curr. Protoc. Hum. Genet. 56:2.10.1-2.10.8. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • genotyping;
  • TaqMan;
  • SNP;
  • AutoProbeMaker;
  • PCR;
  • fluorescent