Sureselect human all exon v5 lncrna kit

Sequencing libraries were prepared, and captured using SureSelect Human All Exon V5+lncRNA kit (Agilent Technologies, Santa Clara, CA, USA) following the manufacturer’s instructions. In brief, 1 μg of DNA was fragmented using a Covaris S2 system (Covaris Inc. Woburn, MA, USA) to produce fragments with an average size of 150–200 base pairs, followed by end repair, A-tailing, and ligation of SureSelect adapter oligos using SureSelect XT Reagents Kit (Agilent Technologies). Precapture polymerase chain reaction (PCR) amplification of the adapter-ligated library was performed for 5–6 cycles. Next, 750 ng of the amplified libraries were hybridized with the SureSelect capture library for 24 h and purified. Post-capture amplification was performed for 10–11 cycles using Herculase II polymerase with SureSelect Indexing Post-Capture PCR primers. Purification was performed with Agencourt AMPure XP beads (Beckman Coulter, Brea, CA, USA). Quantification and size distribution of the amplified libraries were determined using a BioAnalyzer (Agilent Technologies) and KAPA Library Quantification Kit (KAPA Biosystems).

Whole-exome capture libraries were prepared from tumor and matched normal DNA, using a SureSelect Human All Exon V5+lncRNA kit (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s instructions. DNA libraries were sequenced in a paired-end mode, using HiSeq 2000 platform (Illumina Inc., San Diego, CA, USA).