WES of PMC42-LA
Exome sequencing libraries were constructed by following manufacturers protocol (SureSelectXT Human All Exome V6/V6 + UTRs kit). For each sample, 100~200ng genomic DNA was randomly fragmented to <300 bp by sonication (Covaris S220). The fragments were treated with End Prep Enzyme Mix for end repairing, then their sizes were selected by AxyPrep Mag PCR Clean-up (Axygen). Poly-A tails were added to 3 ends by A Tailing Mix, and then purified by AxyPrep Mag PCR Clean-up. Next, adaptors were ligated to both ends of purified results. After one more AxyPrep Mag PCR Clean-up size selection, the results were amplified by using Pre-Capture PCR primers. The PCR products were validated by using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA), and quantified by Qubit3.0. To capture exon regions, Block Mix, Hybridization Buffer and Capture Library were poured in 750ng library for 24h. Dynabeads MyOne Streptavidin T1 magnetic beads was used to elute the capture product. Each sample was then amplified by PCR P5 and P7 primers (P7 primer containing 8bp index for further sample identification), At last, Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and Qubit3.0 were used to detect quality and concentration of libraries, respectively.
SUB6087205
PMC42-LA WES