GSM6765200: ATACseq_R1mESC_1; Mus musculus; ATAC-seq
Nuclei were isolated from cryopreserved cells. Per replicate, roughly 50,000 nuclei were tagmented for 5 minutes with in-house generated Tn5 while shaking at 37°C and DNA was purified using 2x SPRI beads (Ampure) purification. DNA was subsequently PCR amplified by using KAPA HiFi Hotstart Ready Mix (KAPA Biosystems) and Nextera Index Kit (Illumina) primers followed by reverse phase 0.65 x SPRI beads purification and a column purification (Zymo). To remove large DNA fragments from the libraries, samples were subjected to another 0.5X SPRI beads purification, followed by a 1X purification. Samples were analyzed for purity using a High Sensitivity DNA Chip on a Bioanalyzer 2100 system (Agilent). Libraries were paired-end sequenced on a Illumina NextSeq500.