GSM7506230: BDL KO2; Mus musculus; ATAC-seq
Mouse tissue samples were first homegenised to obtain intact nuclei and then treated with a highly active Tn5 transposable mutant capable of simultaneously tagging the target DNA with a sequencing adapter, thus completing the fragmentation of the DNA also called “tagmentation". Labeling and library preparation were performed using the ATAC-Seq Kit (Catalog No. 53150 Active Motif North America) according to the instructions provided within the kit. The nuclei were first counted and centrifuged at 4°C, 500g speed for 5 min to prepare aliquots containing 50,000 to 100,000 nuclei. The samples were resuscitated with 100 μl ice-cold ATAC Lysis Buffer and to each sample was added 50 μl Tagmentation Master Mix and incubated in a thermomixer at 800 rpm and 37℃ for 30 minutes. The library was then prepared by PCR amplification of the labelled DNA using a unique combination of Illumina's i7/i5 primers based on a Nextera adapter. Finally, the library was purified using SPRI bead washing. Two biological replicates per condition were performed.