title: GSM4041816: DNMT3A/DNMT3B-DKO; Homo sapiens; Bisulfite-Seq
library_construction_protocol: To prepare genomic DNA, cells were resuspended with cell lysis buffer (10 mM Tris-HCl pH 7.5, 10 mM EDTA, 10 mM NaCl, 0.5% sarcosyl, 0.1 mg/mL RNase (CWBIO)) and incubated at 37 °C overnight. Then, protease K (Merck) was added to a final concentration of 0.2 mg/mL and incubated at 65 °C for 24 hr. Genomic DNA was extracted by phenol/chloroform and ethanol precipitated. The DNA was dissolve in ddH2O. The purified genomic DNA was digested overnight with MspI (NEB). The sticky ends produced by MspI digestion were filled with CG nucleotides, and 3′A overhangs were added. Methylated Illumina sequencing adapters with 3′T overhangs were ligated to the digested DNA. 40–220 bp insert sizes were converted by bisulfite using an EZ DNA Methylation Gold kit (Zymo Research, USA). The libraries were PCR amplified and each library was sequenced by an Illumina HiSeq platform PE150 (150 bp paired-end reads).
attributes.tag: GEO Accession
attributes.value: GSM4041816