title: GSM4193088: Myotubes_MeDIP; Mus musculus; MeDIP-Seq
library_construction_protocol: Lysates were clarified from sonicated nuclei In brief, DNA libraries were prepared with the NEB Next DNA Library Prep Master Mix Set for Illumina (E6040, NEB) following the manual provided by the manufacturer.First, fragmented DNA (200-500 bp) was end repaired and purification by AMPure XP Beads (A6380, Beckman). Then, purified DNA was added dA-tails and purified with AM-Pure XP Beads again. Subsequently, dA tailed DNA was ligated with adaptors. After purifi-cation, treated DNA was denatured at 99°C for 10 min, then placed the tube on ice immedi-ately. Next, DNA was incubated with the specific antibody against 5mC (BI-MECY-0100) at 4°C for 2 h and captured with protein A/G agarose beads at 4 °C for 2 h. After washing three times with IP buffer (140 mM NaCl, 0.05% Triton X-100, 3.9 mM NaH2PO4, 6.1 mM Na2HPO4) and digesting with proteinase K, the DNA fragments were extracted twice by phenol-chloroform and amplified by adaptor-mediated PCR. Subsequently, after purifica-tion, the DNA fragments were sequenced by the Illumina HiSeq 2000 sequencing platform.
attributes.tag: GEO Accession
attributes.value: GSM4193088