Mung bean nuclease treatment increases capture specificity of microdroplet-PCR based targeted DNA enrichment.

ABSTRACT: Targeted DNA enrichment coupled with next generation sequencing has been increasingly used for interrogation of select sub-genomic regions at high depth of coverage in a cost effective manner. Specificity measured by on-target efficiency is a key performance metric for target enrichment. Non-specific capture leads to off-target reads, resulting in waste of sequencing throughput on irrelevant regions. Microdroplet-PCR allows simultaneous amplification of up to thousands of regions in the genome and is among the most commonly used strategies for target enrichment. Here we show that carryover of single-stranded template genomic DNA from microdroplet-PCR constitutes a major contributing factor for off-target reads in the resultant libraries. Moreover, treatment of microdroplet-PCR enrichment p

SUBMITTER: Yu Z 

PROVIDER: S-EPMC4110027 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature