title: GSM1621441: RRBS_H2O2_HEK293T; Homo sapiens; Bisulfite-Seq
library_construction_protocol: Total DNA of HEK293T cell line with or without H2O2 treatment was extracted by proteinase K/phenol. For each library, 500 ng of genomic DNA was digested overnight with MspI (20 units). After purification, the digested products were blunt-ended. Then dA was added, followed by methylated-adapter (Illumina, CA, USA) ligation. To obtain DNA fractions in 40-120 bp and 120-220 bp range of MspI-digested products, two ranges (160-240 bp and 240-340 bp) of adapter-ligated fractions were excised from 2% agarose gel. The size-selected DNA was bisulfite-treated for two rounds using EpiTect Bisulfite Kit (Qiagen, Venlo, Netherlands) following the manufacturer's standard protocol. Finally, libraries were generated by amplification with HiFi HotStart Uracil+ ReadyMix (KAPA, MA，USA) and Illumina Multiplexing PCR Primers. Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA) and RT-PCR were used to guarantee the quality and quantity of RRBS library. After RRBS library construction, Illumina HiSeq 2000 was used to sequence those libraries according to the manufacturer's protocol.
attributes.tag: GEO Accession
attributes.value: GSM1621441