GSM2308832: 6h embryo DMAD knock down 6mA MeDIP-seq; Drosophila melanogaster; MeDIP-Seq
Genomic DNA was extracted from Drosophila and other animal tissues using the Wizard genomic DNA purification Kit (Promega) following the manufacturer's instructions. 5 µg of fragmented genomic DNA was immunoprecipitated with 5 µg of rabbit 6mA antibody (Abcam) or rabbit IgG overnight at 4°C in a final volume of 500 µl immunoprecipitation buffer. Then the mixture was incubated with 50 µl protein A/G agarose beads for 2 h at 4°C and then washed three times with 1 ml of immunoprecipitation buffer. The beads were then treated with proteinase K and the methylated DNA was purified by phenol-chloroform extraction followed by ethanol precipitation for library construction. End repair, 3'-adenylation, adaptor ligation, and PCR amplification were performed according to Illumina TruSeq DNA sample preparation procedures. Libraries were sequenced using the Illumina Hiseq2500 V4 platform.