title: GSM2816570: KO_5; Mus musculus; ATAC-seq
library_construction_protocol: Adipocyte nuclei were isolated according to methods previously described (Church 2014). Briefly, 250,000 cells from filtered adipocyte suspension were washed once with PBS prior to addition of cold hypotonic lysis buffer containing 0.1% Igepal to yield nuclei. For SVF, 50,000 cells were used for nuclei isolation according to standard protocol. The lysis reaction was centifuged and the supernatant was discarded. Isolated nuclei were incubated with 10x concentration Tn5 (Nextera) for primary adipose and standard concentration for SVF. 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: GSM2816570