Estimation of single nucleotide polymorphism allele frequency in DNA pools by using Pyrosequencing.

摘要: Positional cloning of genes underlying complex diseases, such as type 2 diabetes mellitus (T2DM), typically follows a two-tiered process in which chromosomal region is first identified by genome-wide linkage scanning, followed association analyses using densely spaced single nucleotide polymorphic markers to identify the causal variant(s). The success polymorphism (SNP) detection has resulted vast number potential available for use construction dense SNP maps. However, cost genotyping large numbers SNPs appropriately sized samples nearly prohibitive. We have explored pooled DNA means identifying differences allele frequency between pools individuals with T2DM and unaffected controls Pyrosequencing technology. found that frequencies were strongly correlated those (r=0.99, P<0.0001) across wide range (0.02–0.50). further investigated sensitivity this method detect contrived pools, also over frequencies. was able an difference less than 2% indicating may be sensitive enough studies involving diseases where small cases expected.