title: GSM5515105: OCIAML2 Runx1wt/wt Control ATAC replicate 1; Homo sapiens; ATAC-seq
library_construction_protocol: AML cells (75,000 cells) were treated with nuclear isolation buffer and centrifuged for 10 minutes. Following this, mutant Tn5 transposase was added to the nuclei preparation. Tagmentation reactions were carried out at 37°C for 30 minutes to fragment the accessible genomic DNA and simultaneously add sequencing adapter sequences. Tagmentation reactions were stopped by the addition of binding buffer from a PCR purification kit and the tagmented DNA fragments were purified over a column. The eluted DNA fragments were utilized for library preparation and amplification. PCR Amplified libraries were size selected with AMPure XP beads to remove unbound Illumina index primers. Finished libraries were pooled and paired-end sequenced on a NextSeq 500 sequencer with a 150 cycle mid-output kit. ATAC-Seq Libraries were prepared according to Illumina's instructions accompanying the Nextera DNA Library Prep Kit (Cat. No. FC-121-1030). After adapter ligation, DNA was PCR amplified with Illumina universal primer for 12-15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were isolated utilizing AMP-XP beads. The purified DNA was captured on an Illumina NExt-Seq 500 flow cell for cluster generation. Libraries were sequenced following the manufacturer's protocols.