title: GSM4142786: T47D ATAC-seq noSerum replicate2; Homo sapiens; ATAC-seq
library_construction_protocol: For nuclei preparation, 5x10e6 cells were washed with cold phosphate-buffered saline (PBS), harvested in cold PBS supplemented with protease inhibitor cocktail (PIC, Roche), and cell pellets obtained by centrifugation at 900xg for 5 min at 4ºC. The cell pellet was gently resuspended in 50 µl of RBS buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2) supplemented with PIC, and then 1.3 ml of RBS buffer - 0.1% Igepal CA-630 was added, followed by centrifugation at 500xg for 10 min at 4ºC. The supernatant was carefully discarded and 1 ml of RBS buffer was added to resuspend the cell nuclei. For the transposition reaction, 50,000 nuclei were resuspended in 25 µl 2X TD Buffer (Illumina 121-1030), 2.5 µl Tn5 Transposase (Illumina 121-1030) in a final volume of 50 µl. The reaction mix was incubated for 30 min at 37ºC C and DNA purified using the Qiagen Mini-Elute Kit in 10 µl Tris buffer 10mM, pH8.0. Eluate was subjected to PCR amplification in a volume of 50 µl as follow
attributes.tag: GEO Accession
attributes.value: GSM4142786