title: GSM2559687: WT_dmso4; Mus musculus; ATAC-seq
library_construction_protocol: Primary peritoneal macrophages were isolated four days after thioglycollate injection. Cells were treated for 3 hours and 50,000 cells were centrifuged and washed once with PBS prior to addition of 50uL cold hypotonic lysis buffer containing 0.1% Igepal to yield nuclei. The lysis reaction was centifuged and the supernatant was discarded. Isolated nuclei were incubated with 2.5uL Tn5 (Nextera) in a 50uL reaction volume and placed in a 37C thermomixer for 30 minutes at 300rpm agitation. All ATAC datasets were generated using Tn5 from Nextera with previously published protocols with minor adjustments (Buenrostro et al, Curr.Protoc. Mol. Biol. 2015). Transposed DNA was cleaned up with a Qiagen MinElute PCR Purification Kit and amplified with a non-saturating PCR to enrich for Transposed DNA. The reaction was size selected on a gel for fragments 100-1000bp, were quantified with qPCR and Qubit, and sequenced 50bp, single-end sequencing.
attributes.tag: GEO Accession
attributes.value: GSM2559687