alias: GSM3376455
title: GSM3376455: ATAC-seq_Melanoma_p44_STAT3_KO_113-1_1; Mus musculus; ATAC-seq
library_construction_protocol: Duplicates of two different STAT3fl and STAT3∆ cell lines were used for the assay. A volume containing 50.000 cells were harvested with the aid of an automated cell counter (TC20 Bio-Rad) and spun at 500g for 5 minutes at 4°C in PBS. After centrifugation, the pellet was carefully resuspended in the transposase reaction mix (12.5 µl 2xTD buffer, 2 µl TDE1 (Illumina), and 10.25 µl nuclease-free water, 0.25 µl 1% Digitonin (Sigma)) for 30 min at 37 °C. Following DNA purification with the MinElute kit eluting in 12 µl, 1 µl of the eluted DNA was used in a quantitative PCR reaction to estimate the optimum number of amplification cycles. Library amplification was followed by SPRI size selection to exclude fragments larger than 1,200 bp. DNA and concentration was measured with a Qubit fluorometer (Life Technologies). Library amplification was performed using custom Nextera primers.
identifiers.namespace: GEO
identifiers.id: GSM3376455