title: ovarian cells in culture
design: Exome interrogation was performed using the Illumina TruSeq Exome Library Prep Kit (Illumina FC-150-1001). High Molecular Weight (HMW) genomic DNA (gDNA) was fragmented using the Covaris S2 Ultra-sonication system. Following fragmentation, DNA is end repaired and 3 adenylated prior to adaptor ligation using TruSeq DNA indexed adapters. The four DNA libraries were equalized to 233ng and pooled for two rounds of enrichment using the TruSeq Exome capture probes. The TruSeq Exome capture probes target 45Mb of coding sequence from >98% of RefSeq, CCDS and Ensembl coding content. Following the final exome capture the enriched libraries are PCR amplified to generate sufficient yield for sequencing. Enriched, amplified libraries were validated using the Agilent Bioanalyzer 2100 system and quantitated using the Roche LightCycler96 RealTime PCR system. Final library concentration results were used to dilute the library pool to 2nM in preparation for cluster generation. The cluster generation for two Rapid Read flow cells was performed using HiSeq Rapid PE Cluster v2 kits (Illumina PE-402-4002) on the Illumina cBot Cluster Generation Station. The clustered exome libraries were sequenced on the Illumina HiSeq 2500 sequencer using the TruSeq Rapid SBS Kit v2 200 cycle kits (Illumina FC-402-4021). Following sequence data collection on the Illumina Compute server (iCompute), the raw base call files (.bcl) are converted to fastq files and de-multiplexed into individual libraries.