Targeted-Capture of homo sapiens: adult glioblastoma tumors
DNA was extracted from tissue using the Qiagen?DNeasy?Blood & Tissue Kit (Qiagen 69504). Additional purification was performed using Agencourt AMPure XP beads (Beckman Coulter A63881). DNA quantity was measured using?Quant-iT?PicoGreen?dsDNA Assay kit?(Invitrogen?P7589).? DNA integrity was assessed using an Agilent?TapeStation?with Genomic DNA?Screentape?and reagent?(Agilent 5067-5365,?5067-5366).? DNA libraries were generated with?the Kapa?HyperPlus?Kit?(KK8514)?according to standard protocols?followed by?AMPure?XP bead purification. The library quantity was measured with?picogreen?and quality was verified with a bioanalyzer (Agilent, DNA 1000 chip).?The?University of Texas Southwestern Medical Centers custom?SeqCap?EZ Developer Probe kit 1425?gene panel (Roche NimbleGen REF 06471722001, IRN 4000033290, GTIN 07613336140413)?was used for target hybridization?with?X Gen Universal Blocking Oligo (Integrated DNA Technologies). After hybrid capture,?samples were PCR?amplified, bead purified,?and?quality?checked?using?picogreen?and?bioanalyzer,?then?adapter ligation efficiency?was?measured?by qPCR.?The samples were sequenced on an Illumina?NextSeq?550 sequencer (SY-415-1002) using 75-bp paired-end protocol.?