GSM4629202: EP_DMC1_ChIPSeq Rep 1; Mus musculus; ChIP-Seq
Testes were decapsulated, fixed with 1% formaldehyde, and homogenized to isolate germ cells. Cells were lysed, and chromatin was fragmented by sonication. Cross-links were reversed and chromatin eluted by incubation at 65 degrees Celsius. After high-speed spin soluble chromatin was removed and used for ChIP with anti-DMC1 antibody. For library preparation, DNA was first end-repaired by incubation with 1 × T4 DNA Ligase Reaction Buffer with 0.25 mM dNTP, 3 units of T4 DNA polymerase (NEB, M0203S), 1 unit of Klenow Enzyme (NEB, M0210S) and 10 units of T4 PNK (NEB, B0202S) at 20°C for 30 min, followed by addition of 3′-A overhangs using Klenow Fragment 3′-5′ exo- (NEB, M0212S). After denaturation of DNA at 95°C for 3 min, adapters from the TruSeq Nano DNA LD Library Prep Kit (set A, Illumina, FC-121-4001) were ligated with a Quick Ligation kit (NEB, M2200S). The libraries then were amplified using PCR Enhancer mix and primer cocktail (Illumina, FC-121-4001) for 12 cycles. For each step, DNA was purified by MinElute Reaction Cleanup Kit.
GEO Accession
GSM4629202