title: GSM9076827: SK-N-SH cells expressing dox-inducible WWTR1, treatment with doxycycline; Homo sapiens; ATAC-seq
library_name: GSM9076827
library_construction_protocol: 50 000 cells were detached and dissociated using accutase and washed 1x with DPBS. Cell pellets were lysed in 25 µl Tagment DNA Enzyme and Buffer (Illumina), supplemented with 0.01 % digitonin (Promega) and 2x cOmplete, EDTA-free protease inhibitor cocktail (Roche). Samples were incubated at 37 °C for 30 min before placing on ice and subsequent purification using MinElute PCR purification kit (Qiagen) according to manufacturer's instructions, with elution in 12 µl. 1 μl of eluted DNA was used in a quantitative PCR (qPCR) reaction to estimate the optimum number of amplification cycles. The remaining 10 μl of each library were amplified for the number of cycles corresponding to the Cq value (that is, the cycle number at which fluorescence has increased above background levels) from the qPCR. Library amplification was followed by SPRI (Beckman Coulter) size selection to exclude fragments larger than 1,200 bp. DNA concentration was measured with a Qubit fluorometer (Life Technologies). Library amplification was performed using custom Nextera primers.