title: GSM7683915: INO80-OE adult cardiomyocyte, tamoxifen treatment 2 days, rep2; Mus musculus; ATAC-seq
library_construction_protocol: Aliquots of 50,000 cardiomyocytes were lysed with 3 ml chilled lysis buffer (3.75 mmol/L PIPES, 450 mmol/L KCl, 1% NP-40, 1% Tween-20, 1% Triton X-100 in water; pH 7.3) for 6 minutes, immediately spun down at 500 ´ g for 10 minutes at 4°C. Nuclear pellets were transposed with 25 µl Tagment DNA Buffer, 2.5 µl Tagment DNA Enzyme (Nextera Sample Prep Kit from Illumina, FC-121-1030), 16.5 µl PBS, 0.5 µl 1% digitonin, 0.5 µl 10% Tween-20, and 5 µl nuclease-free water. Samples were mixed and incubated at 37°C for 30 minutes in a thermomixer at 1000 rpm. Transposed samples were purified with the QIAGEN MinElute Reaction Cleanup Kit (28204) and amplified with 25 µl NEB Next High-Fidelity 2× PCR Master Mix (M0541S), 1.25 mmol/L Nextera custom primers with unique barcodes, and nuclease-free water. Samples were amplified at the following PCR conditions: 72 °C for 5 min and 98 °C for 30 sec and cycled at 98 °C for 10 sec, 63 °C for 30 sec, and 72 °C for 1 min. Half of each sample was amplified for 12 cycles, purified, and assessed by Bioanalyzer (Agilent) for library quality. Sample concentration was quantified by Qubit (Invitrogen) before pooling. Pooled samples were sequenced with a PE75 run on a NextSeq500 instrument (Illumina).