GSM4816108: adipocyte Dnmt1 AKO (Bisulfite-Seq); Mus musculus; Bisulfite-Seq
Briefly, primary adipocytes were lysed for 1 h in lysis buffer at 37°C and digested with 10 g/mL proteinase K for 3 h at 50°C. After cell lysis, DNA and RNA was isolated by phenol-chloroform extraction and using trizol, respectively. For WGBS library construction, 100ng of genomic DNA was treated with the EZ DNA Methylation-Gold kit (Zymo Research, Orange, CA, USA) for bisulfite conversion. Then, the sequencing libraries were prepared according to the manufacturer's instructions of TruSeq DNA Methylation Library Preparation Kits (Illumina, #EGMK91396). Briefly, bisulfite-converted DNA is random-primed using a polymerase to synthesize DNA containing a specific sequence tag. The 3' ends of the newly synthesized DNA strands are tagged with a second specific sequence. The di-tagged DNA molecules are amplified using FailSafe PCR Enzyme (Epicentre, #FSE51100) and TruSeq Index PCR Primer. The final purified product is quantified using qPCR according to the qPCR Quantification Protocol Guide and qualified using the Agilent Technologies 2100 Bioanalyzer (Agilent). Finally, sequencing was performed using the HiSeqX platform (Illumina). In case of RNA-seq,   RNA libraries were prepared for sequencing using standard Illumina protocols.
GEO Accession
GSM4816108