title: GSM3465058: Histone_H1_ChIP_OE_rep2; Mus musculus; ChIP-Seq
library_construction_protocol: Cross-linking was done by incubating MOE cells with 1% formaldehyde for 5 minutes. After washing off the formaldehyde, MOE cells were lysed briefly in SDS containing buffer followed by sonication for 15 minutes with 30 second intervals to shear genomic DNA using Bioruptor UCD200 (Diagenode). The sheared DNA was evaluated by electrophoresis for their size ranging between 200 bp and 300 bp. Each sheared genomic DNA preparation was split into two equal portions and incubated with either MeCP2 (Diagenode, pAb-052-050) or Histone H1 (Santa Cruz, sc-34464) or H3K27me3 (Diagenode, pAb-069-050), H3K9ac (Diagenode, pAb-177-050) or Rabbit IgG (Millipore, 12-370) for the negative control. A magnetic bar was used to pull down antibody corresponding protein-genomic DNA complex. Genomic DNA in the complex was reverse cross-linked by DNase-free Proteinase K and purified. Histone H1, H3K27me3, and H3K9ac ChIPed or input DNA were used to prepare the ChIP-Seq library as described (NEXTflex ChIP-Seq kit, Bio Scientific, NOVA 5143-01 and NEXTflex ChIP-Seq Barcodes-6, NOVA 514120) followed by high throughput sequencing with Illumina Hi-seq.2000
attributes.tag: GEO Accession
attributes.value: GSM3465058