GSM3593925: ssDIP; Mus musculus; MeDIP-Seq
Cellular genomic DNA was purified using the DNeasy kit (QIAGEN, 69504) with minor modifications. In brief, the cell lysate was treated with RNAse A1/T mix at 37 °C for 30 min. Then Proteinase K was added to the lysate and treated at 56 °C for 3 hours or overnight. The remaining steps were performed as described in the kit, and the DNA was eluted with ddH2O. In briefly, 5 μg extracted genomic DNA was sonicated to 200–500 bp with Bioruptor. Then, adaptors were ligated to genomic DNA fragments following the Illumina protocol. The ligated DNA fragments were denatured at 95 °C for 5 min. Then, the single-stranded DNA fragments were immunoprecipitated with N6-mA antibodies (5 μg for each reaction, 202-003, Synaptic Systems) overnight at 4 °C. N6-mA enriched DNA fragments were purified according to the Active Motif hMeDIP protocol. IP DNA and input DNA were PCR amplified with NEBNext Multiplex Oligos for Illumina indexing primers.
GEO Accession
GSM3593925