Double indexing overcomes inaccuracies in multiplex sequencing on the Illumina platform.

ABSTRACT: Due to the increasing throughput of current DNA sequencing instruments, sample multiplexing is necessary for making economical use of available sequencing capacities. A widely used multiplexing strategy for the Illumina Genome Analyzer utilizes sample-specific indexes, which are embedded in one of the library adapters. However, this and similar multiplex approaches come with a risk of sample misidentification. By introducing indexes into both library adapters (double indexing), we have developed a method that reveals the rate of sample misidentification within current multiplex sequencing experiments. With ~0.3% these rates are orders of magnitude higher than expected and may severely confound applications in cancer genomics and other fields requiring accurate detection of rare variants. W

SUBMITTER: Kircher M 

PROVIDER: S-EPMC3245947 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature