GSM1981827: BGA81; Bos taurus; Bisulfite-Seq
Genomic DNA for each tissue was isolated according to the protocol of QIAamp@ DNA Mini Kit (QIAGEN). RRBS libraries were constructed according to the manufacturer’s instruction. In detail, 3ug genomic DNA was digested with the methyl insensitive MspI enzyme (CCGG site) at 37 ºC for 16h for each samples, individually. The digested DNA products were purified using QIAquick PCR Purification Kit (QIAGEN) and added single A nucleotides in the blunt-end, which were then ligated to methylated adapter with T overhang. The DNA fragments with 150-400bp length were collected and purified using 2.5% agarose gel electrophoresis. The recovered DNA was treated with methylation-Gold Kit (ZYMO) to apply for the bisulfite conversion. The bisulfite-converted DNA was finally amplified by PCR to construct the RRBS libraries. The Agilent 2100 bioanalyzer instrument (Agilent DNA 1000 Reagents) and the real-time quantitative PCR (qPCR, TaqMan Probe) were used to quantitate and quantify the RRBS libraries, separately. The qualified libraries were amplified on cBot to generate the cluster on the flowcell (TruSeq PE Cluster Kit V3-cBot-HS, Illumina).
GEO Accession
GSM1981827