Nimblegen seqcap ez exome v2 kit

Whole-genome amplified DNA was fragmented using the Covaris Sonicator to 250 bp and purified using a Zymo DNA Clean & Concentrator Column kit (Zymo, D4004) according to the manufacturer's instructions. Barcoded next-generation sequencing libraries were constructed using the NEBNext end repair model (NEB, E6050L), dA-tailing module (NEB, E6053L), and quick ligation module (NEB, E6056L). Libraries were amplified via PCR using NEBNext HiFi2x PCRmix (NEB, M0541L). Targeted capture for single cells was performed using Nimblegen SeqCap EZ Choice Library, according to Leung et al. (2016) (link). Exome capture for CRC1 population was performed on single-cell sequencing libraries using the TruSeq Exome Enrichment kit (Illumina, 15013230) following the manufacturer's instructions. Exome capture for CRC2 population was performed using a Nimblegen SeqCap EZ Exome V2 kit (Roche, 05860482001). For exome or targeted-capture sequencing, samples were sequenced on a 100 pair-end flow-cell on the Illumina HiSeq 4000 system. For copy number profiling, barcoded libraries were pooled using equimolar concentrations and sequenced at 76 single-read flow-cell on the Illumina HiSeq 2000 system.