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. 2008 Jan:Chapter 2:Unit 2.10.
doi: 10.1002/0471142905.hg0210s56.

Genotyping using the TaqMan assay

Lester Hui  1 , Terrye DelMonte, Koustubh Ranade
Affiliations

Genotyping using the TaqMan assay

Lester Hui et al. Curr Protoc Hum Genet. 2008 Jan.

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.

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