title: GSM4259843: Runx1Evi1_NoDox_HE_DHS; Mus musculus; OTHER
library_construction_protocol: DNaseI was performed as previously described [Bert et al., 2007. PMID:17283044]. 3x10^5 sorted cells were added directly to DNaseI (Worthington Biochemical Corporation) used between 6 and 13 U/ml for 3 minutes at 22°C. The reaction was terminated by addition of SDS to 0.5% and cell lysates treated with 0.5 mg/ml proteinase K overnight at 45°C overnight then 0.2 mg/ml RNaseA for 1 hour at 37°C. DNA was isolated by phenol/chloroform extraction and level of DNaseI digestion was assessed using qRT-PCR for the active TBP promoter compared to an inactive region of chromosome 1 to ensure comparable digestion points were used. DNaseI fragments were size selected on an agarose gel to between 50 and 250 bp then used to generate a library using the KAPA hyper prep kit, according to the manufacturer's instructions.  These were then further size selected to obtain fragments between 200 and 300 bp and sequenced single-end in a pool of 12 indexed libraries using a Next-Seq 500 high output kit v2 75 cycles (Illumina) at the Genomics Birmingham sequencing facility.
attributes.tag: GEO Accession
attributes.value: GSM4259843