GSM5183598: Methyl caputre seq #2 on H1 QSER1/DNMT3B DKO hESCs; Homo sapiens; Bisulfite-Seq
DNA was extracted using the DNeasy Blood & Tissue Kit (Qiagen, 69504) following manufacturer's guidelines. Two biological replicates for each genotype were submitted to the MSKCC Integrated Genomics Operation Core for library prep and sequencing. After PicoGreen quantification and quality control by Agilent BioAnalyzer, 480-490ng of genomic DNA were sheared using a LE220-plus Focused-ultrasonicator (Covaris catalog # 500569). Sequencing libraries were prepared using the KAPA Hyper Prep Kit (Kapa Biosystems KK8504) without PCR amplification. Post-ligation cleanup proceeded according to Illumina's instructions with 110 µL Sample Purification Mix from the TruSeq Methyl Capture EPIC LT Library Prep Kit (Illumina catalog # FC-151-1002). After purification, 4 samples were pooled equimolar and methylome regions were captured using EPIC oligos. Capture pools were bisulfite converted and amplified with 11 cycles of PCR. Pools were sequenced on a HiSeq 4000 in a 100bp/100bp paired end run, using the HiSeq 3000/4000 SBS Kit (Illumina). The average number of read pairs per sample was 29 million. TruSeq Methyl Capture EPIC Library Prep Kit from Illumina
GEO Accession
GSM5183598