GSM1876032: Athero_Artery_ATAC_D4; Homo sapiens; OTHER
10-20 mg of pulverized fresh frozen tissue was suspended in 1 mL 1x PBS and centrifuged at 2,000 x g for 3 min at 4° C. The pellet was resuspended in 1 mL of a lysis buffer containing 50 mM HEPES, pH 7.5, 140 mM NaCl, 1 mM EDTA, 10% glycerol, 0.5% IGEPAL CA-630, and 0.25% Triton X-100 and supplemented with an EDTA-free complete protease inhibitor (Roche). This suspension was incubated at 4° C for 10 minutes and then dounce homogenized. The homogenate was centrifuged at 2,000 x g for 5 min at 4° C, and the resultant pellet was resuspended in 1 mL 1x PBS. Cellular debris was filtered with a cell strainer, and intact nuclei were quantified. 5x10^4 nuclei were subjected to Tn5 mediated transposition for 1 hour and tagmented DNA was purified as previously reported. Libraries were PCR amplified and purified as described previously and were subjected to an additional electrophoresis step on an E-Gel EX 2% agarose gel (Invitrogen), and gel fragments correlating to 100-1000 bp were excised and purified. The quality of the library preparation was determined by evaluation of the electropherogram traces from an Agilent 2100 Bioanalyzer DNA High Sensitivity, and libraries demonstrating appropriate nucleosomal enrichment were multiplexed. Libraries were quantitated using Qubit high sensitivity DNA reagent, and quantitative PCR (KAPA Biosystems).
GEO Accession
GSM1876032