title: GSM8491330: U87MG cells, IDH1 WT expression, tumor collected 27 days after cell injection, sample 21; Mus musculus; Bisulfite-Seq
library_construction_protocol: 100–200 mg of tumor tissue from 22 U87MG and HT1080 xenografts tumors was weighed and then shipped in dry ice for RRBS analysis at Active Motif (Carlsbad, CA). There, tissues were subjected to proteinase K digest solution (0.5% SDS, 0.5mg/ml proteinase K, 100 mM EDTA, in TE pH 8) by allowing the samples to rotate overnight at 55⁰C. For library preparation and sequencing, 100 ng of gDNA was digested with TaqaI at 65 ⁰C for 2 h followed by MspI treatment at 37⁰C overnight. Following enzymatic digestion, samples were used for library generation using the Ovation RRBS Methyl-Seq System following the manufacturer's instructions. The digested DNA was randomly ligated, and, following fragment end repair, bisulfite was converted using the EpiTect Fast DNA Bisulfite Kit following the Qiagen protocol. After conversion and clean-up, samples were amplified, resuming the Ovation RRBS Methyl-Seq System protocol for library amplification and purification. Libraries were measured using the Agilent 2200 TapeStation System and quantified using the KAPA Library Quant Kit ABI Prism qPCR Mix. Libraries were sequenced on a NovaSeq 6000 at SE75.