GSM2974475: HCC1_3DPDX_WES; Homo sapiens; OTHER
The quality and quantity of purified DNA were assessed by fluorometry (Qubit, Invitrogen) and gel electrophoresis. Briefly, 500ng of genomic DNA from each samples were fragmented by acoustic shearing on a Covaris S2 instrument. Fragments of 150–300 bp were ligated to Illumina's adapters and PCR-amplified. The samples were concentrated to 300 ng in 3.4 μl DW using a Speedvac machine (Thermo Scientific) and hybridized with RNA probes, SureSelect XT Human All Exon V5 Capture library for 16-24 h at 65°C. After hybridization, the captured targets were pulled down by biotinylated probe/target hybrids using streptavidin-coated magnetic beads (Dynabeads My One Streptavidine T1; Life Technologies Ltd.) and buffers. The selected regions were then PCR-amplified using Illumina PCR primers. Libraries were quantified using the Agilent 2100 Bioanalyzer (Agilent Technologies) and KAPA Library Quantification Kit (KK4824, Kapa Biosystems). The resulting purified libraries were applied to an Illumina flow cell for cluster generation and sequenced using 100 bp paired-end reads on an Illumina Hiseq2500 sequencer by following the manufacturer's protocols. Image analysis were performed using the HiSeq control Software version 1.8.4.
GEO
GSM2974475