title: GSM3018360: pol II ChIPseq 0hr control; Mus musculus; ChIP-Seq
library_construction_protocol: Cell pellets were treated with different chromatin extraction buffer supplemented with protease and phosphatase inhibitors (Roche)  then clarified from sonicated nuclei and histone-DNA complexes were isolated with respective antibodies. 50 µl blocked protien A sepharose beads were added to each sample and incubated for 2.5h at 4°C to pull down the respective antibody-chromatin complexes. The beads were then washed with different washing buffers. Protein-bound chromatin complexes were eluted from beads for 30 min using elution buffer followed by reverse-crosslinked by incubating the eluted supernatant at 65°C overnight. Next day DNA was purified from the reverse cross-linked chromatin by proteinase-K and RNase digestion followed by purification using PCR purification kit (Qiagen). For ChIP-seq library preparation, 30 µl ChIP-DNA was processed for library preparation according to ChIP-seq library preparation recommended protocol (NEB)
attributes.tag: GEO Accession
attributes.value: GSM3018360