title: whole exome-seq of homo sapiens: adult  blood
design: For the generation of standard exome capture libraries, we used the Agilent SureSelect Target Enrichment protocol for Illumina paired-end sequencing library (ver. B.3, June 2015) together with 1ug input gDNA. In all cases, the SureSelect Human All Exon V5 probe set was used. The quantification of DNA and the DNA quality is measured by PicoGreen and agarose gel electrophoresis. We used 1g of each cell line's genomic DNA diluted in EB Buffer and sheared to a target peak size of 150200 bp using the Covaris LE220 focused-ultrasonicator (Covaris, Woburn, MA) according to the manufacturer's recommendations. Load the 8 microTUBE Strip  into the tube holder of the ultrasonicator and shear the DNA using the following settings: mode, frequency sweeping; duty cycle, 10%; intensity, 5; cycles per burst, 200; duration, 60 sec  6 cycles; temperature, 4C7C. The fragmented DNA is repaired, an A is ligated to the 3 end, agilnet adapters are then ligated to the fragments. Once ligation had been assessed, the adapter ligated product is PCR amplified. The final purified product is then quantified using the TapeStation DNA screentape D1000 (Agilent). For exome capture, 250 ng of DNA library was mixed with hybridization buffers, blocking mixes, RNase block and 5 l of SureSelect all exon capture library, according to the standard Agilent SureSelect Target Enrichment protocol. Hybridization to the capture baits was conducted at 65C using heated thermal cycler lid option at 105C for 24 hours on PCR machine.  The captured DNA was then washing and amplified. The final purified product is then quantified using qPCR according to the qPCR Quantification Protocol Guide Guide (KAPA Library Quantificatoin kits for Illumina Sequecing platforms) and qualified using the TapeStation DNA screentape D1000 (Agilent). And then we sequenced using the HiSeq 2500 platform (Illumina,San Diego, USA)