alias: DRX001086
title: DCM whole exome sequence id=5
design: none provided
library_construction_protocol: We extracted human genome DNA from 7 ml blood samples using the Wako DNA extraction kit (Wako Chemical, Osaka, Japan) according to the manufacturer?s instructions. The final concentration of DNA solution was adjusted to 500 ng/?L. To prepare fragment libraries for whole-exome resequencing, a total of 1500 ng of each DNA was sheared into 100?150 bp by the Covaris S2 system (Covaris, Woburn, MA, USA) using the following program: 20% duty cycle, 5 intensity, and 200 cycles per burst for 60 seconds at 5?C. Genome DNA was sheared in a 120 ?L reaction in a Covaris microTube containing 10 mM TE buffer. For exome enrichment by hybridization capture, we used an AB SOLiD optimized SureSelect Target Enrichment Human All Exon kit (G3362, Agilent, Santa Clara, CA, USA), which contained exonic sequences of approximately 18000 genes, covering a total of 37.6-megabase genome sequences (approximately 93.8% of all human exons according to the Consensus coding sequences: CCDS, Sept. 2009), following the manufacturer's instructions. The enriched exome libraries were subsequently used for emulsion-PCRs at a library concentration of 0.5 pM following the manufacturer's instructions (Life Technologies). For each of the 8 samples, the beads after emulsion-PCRs were deposited onto one-eighth of a partitioned glass surface of a SOLiD slide. The 50-base sequences were obtained according to the manufacturer?s protocol.
identifiers.namespace: HAMAMATSU
identifiers.id: DRX001086