title: GSM3593926: dsDIP; Mus musculus; MeDIP-Seq
library_construction_protocol: 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.
attributes.tag: GEO Accession
attributes.value: GSM3593926