TP53 R248W mut, glioblastoma
Genomic DNA (gDNA) was isolated from ~25 mg tumor tissue using the QIAamp DNA Mini kit with initial tissue disruption in ATL/proteinase K buffer using a micropestle in Eppendorf Lo-Bind 1.5 mL tubes. Following complete tissue dissolution at 56oC for 2 h, viscous DNA was sheared into uniform 23 kb fragments by passage through a QIAshredder column and centrifugation for 2 min at 15,000 g. The manufacturer's protocol for gDNA purification was subsequently followed for all steps. Libraries were prepared from the purified gDNA at MedGenome for Whole Exome Sequencing using the Agilent SureSelect Human All Exon v7 kit and the manufacturer's protocol. Quality control for the library included Qubit fluorometric quantification, Agilent 4200 tapestation electropherogram, and qPCR.