GSM2701101: 3-4h_dH1; Drosophila melanogaster; ChIP-Seq
1.Nucleosome DNA fragment extraction, chromatin are digested by MNase and nucleosome was dissolved through sonication with 3 cycles of 20s duration with at least 40s pauses between cycles at the power setting of 6 (out of 20) on a Misonix sonicators XL-2000. Then nucleosome DNA was extracted by phenol–chloroform and dissolved in ddH2O. 2. H1 binding DNA fragment extraction, generated chromatin fragments between 100 and 500 bp through sonication with 7 cycles and 5 cycles on 3-4h and 14-15h embryonic nuclei, separately. Then immunoprecipitated DNA-H1 complexes were captured by anti-H1 antibody (Active Motif, 39707) dose as specification described in 20-25ug chromatin.Immunoprecipitated DNAs were obtained by extraction with phenol chloroform after incubated on metal bath under the condition of 65 ℃ over night. Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit.