Allele quantification and DNA pooling methods.

Abstract

Studies utilizing differences in single-nucleotide polymorphism allele frequencies between cases and controls have been widely used in genetic analyses to locate putative genes or chromosomal regions that may be associated with a disease. In these studies the assessment of allele frequencies can be expedited and the genotyping costs reduced by the use of DNA pools. There have been multiple studies that have reported the accuracy of Pyrosequencing for the assessment of allele frequencies in DNA pools. In addition, there are an increasing number of other types of studies that make use of allele quantification to evaluate a disease status or to make a clinical diagnosis. In this chapter, the making of DNA pools is described, as well as the use of Pyrosequencing to quantify alleles. The ease of use, short run, and analysis times make Pyrosequencing the preferred method.