Leveraging a Trans-Omics Strategy for Prioritizing Personalized Candidate Mutation-Driver Genes
Library construction, exome capture, and whole exome sequencing (WES)  The qualified genomic DNA sample was randomly fragmented into fragments with a base pair peak of 150 to 200bp, and then adapters were ligated to both ends of the resulting fragments. The adapter-ligated templates were purified by the AgencourtAMPure SPRI beads and fragments with insert size about 200bp were excised. Extracted DNA was amplified by ligation-mediated (LM-) PCR, purified, and hybridized to the SureSelect Biotinylated RNA Library (BAITS) for enrichment. Hybridized fragments were bound to the streptavidin beads whereas non-hybridized fragments were washed out after 24 hours. Captured LM-PCR products were subjected to Agilent 2100 Bioanalyzer to estimate the magnitude of enrichment. Each captured library was then loaded on Hiseq2000 platform. We performed high-throughput sequencing for each captured library independently to ensure that each sample met the desired average fold-coverage (i.e., at least 100×). Raw image files were processed by Illumina base calling Software (version 1.7) for base calling with default parameters and the sequences of each individual were generated as 90bp paired-end reads.
Chongqing Medical University, Chongqing, China.
the whole exome sequencing (BJ22C_DNA)