title: GSM2259217: D3 BFP Rep2; Mus musculus; OTHER
library_construction_protocol: ATAC-seq was performed according to the published protocol (Buenrostro et al., 2013). For library preparation, 50,000 cells were FACSd into PBS, and then nuclei were isolated by re-suspending the cells in lysis buffer (10mM Tris-HCl ph 7.4, 10mM NaCl, 3mM MgCl2, 0.1% NP-40) for 15 minutes on ice, followed by centrifugation at 500g for 10 minutes at 4oC. Precipitated nuclei were tagmented by re-suspension in the Nextera transposase mix (22.5uL ddH2O, 25uL 2x Tagment DNA buffer, 2.5ul Tagment DNA enyme) (Nextera DNA sample prep kit, FC-121-1030 and FC-121-1011) and incubated at 37oC for 30 minutes. The tagmentation reactions were terminated and cleaned-up using Qiagen Minelute enzyme cleanup kit, eluting in a final volume of 10μl ddH2O. The fragments were amplified by PCR for ~16 cycles, and >100 bp fragments were purified with SeraMag beads.
attributes.tag: GEO Accession
attributes.value: GSM2259217