Variant identification in multi-sample pools by illumina genome analyzer sequencing.

ABSTRACT: Multi-sample pooling and Illumina Genome Analyzer (GA) sequencing allows high throughput sequencing of multiple samples to determine population sequence variation. A preliminary experiment, using the RET proto-oncogene as a model, predicted ≤ 30 samples could be pooled to reliably detect singleton variants without requiring additional confirmation testing. This report used 30 and 50 sample pools to test the hypothesized pooling limit and also to test recent protocol improvements, Illumina GAIIx upgrades, and longer read chemistry. The SequalPrep(TM) method was used to normalize amplicons before pooling. For comparison, a single 'control' sample was run in a different flow cell lane. Data was evaluated by variant read percentages and the subtractive correction method which utilizes the cont

SUBMITTER: Margraf RL 

PROVIDER: S-EPMC3121147 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature