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Clin Chim Acta. 2006 Jan;363(1-2):32-47. Epub 2005 Sep 22.

The use of real-time PCR methods in DNA sequence variation analysis.

Author information

1
R&D Genetics, AstraZeneca Pharmaceuticals, 19G9 Mereside, Macclesfield, Cheshire, UK. neil.gibson@astrazeneca.com

Abstract

BACKGROUND:

Real-time (RT) PCR methods for discovering and genotyping single nucleotide polymorphisms (SNPs) are becoming increasingly important in various fields of biological sciences. SNP genotyping is widely used to perform genetic association studies aimed at characterising the genetic factors underlying inherited traits. The detection and quantification of somatic mutations is an important tool for investigating the genetic causes of tumorigenesis. In infectious disease diagnostics there is an increasing emphasis placed on genotyping variation within the genomes of pathogenic organisms in order to distinguish between strains.

METHODS:

There are several platforms and methods available to the researcher wishing to undertake SNP analysis using real-time PCR methods. These use fluorescent technologies for discriminating between the alternate alleles of a polymorphism. There are several real-time PCR platforms currently on the market. Two of the key technical challenges are allele discrimination and allele quantification.

CONCLUSIONS:

Applications of this technology include SNP genotyping, the sensitive detection of somatic mutations and infectious disease subtyping.

PMID:
16182268
DOI:
10.1016/j.cccn.2005.06.022
[Indexed for MEDLINE]

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