GSM5518404: atac-P-D2; Homo sapiens; ATAC-seq
Viable cells were lysed with digitonin as a detergent and pelleted by centrifugation at 500 RCF for 5 minutes at 4C. The nuclei pellet was resuspended in 50 uL of transposase mixture (25 uL 2x TD buffer, 2.5 uL of TDE1, 16.5 uL PBS, 0.5 uL 1% digitonin,  0.5 uL 10% Tween-20, 5 uL nuclease-free water). Transposition reactions were incubated at 37°C for 30 minutes in an Eppendorf ThermoMixer with agitation at 1000 RPM. Transposed DNA was purified using a Zymo DNA Clean and Concentrator-5 Kit (cat# D4014) and purified DNA was eluted in 20 ul elution buffer (10 mM Tris-HCl, pH 8). Transposed fragments were amplified and purified with modified primers according to Buenrostro 2015. Libraries were quantified using qPCR prior to sequencing. All Fast-ATAC libraries were sequenced using paired-end, dual-index sequencing on an Illumina HiSeq 4000 at a depth of 50 million reads per sample.