ATAC-seq of human: adult male bone_marrow tumor: Sample H_KA-412761-atac2_Lib2
"Transposase-mediated chromatin accessibility profiling of primary AML cells was performed using the Nextera library preparation kit as described in (Buenrostro et al., 2013) using 75,000-150,000 viable cells per sample. Briefly, cells were pelleted, washed once in 50 uL cold PBS, and then resuspended in 50 uL cold ATAC lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Igepal CA-630). Lysed cells were then immediately centrifuged at 500 g for 10 min at 4oC, and cell nuclei were resuspended in 25 uL Nextera tagmentation buffer (Illumina) for transposition with 2.5 uL of Nextera transposase enzyme (Illumina) in a total volume of 50 uL at 37oC for 30 minutes. DNA was purified with Qiagen MinElute columns before amplification with sample-specific primers for a total 9-12 cycles (the exact cycle number was determined via qPCR analysis after 5 cycles). Libraries were either size-fractionated into small (<300 bp) and large (300-800 bp) libraries, or a single library (100-500 bp) "