title: GSM4671367: P3F_high_U23674_ATACSeq; Mus musculus; ATAC-seq
library_construction_protocol: FACS-sorted cells were centrifuged at 500g for 5 min at 4oC. Supernatant was carefully removed and discarded without disturbing the cell pellet. Cell pellets were then resuspended in 50l cold lysis buffer (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% NP40) and immediately centrifuged at 500g for 5 min at 4oC. Supernatant was removed carefully and then resuspended in freshly prepared Tn5 reaction buffer (12.5ml 1X TD buffer (Illumina), 1ml Tn5 Transposase, 11.5ml nuclease-free water). Transposition reaction was incubated at 37oC for 30min and then purified using Zymo ChIP clean up kit. Transposed DNA was subjected to 5 cycles of indexed PCR amplification using NEBNext Ultra II Q5 Master mix and custom indexed primers. Quantitative real time PCR was performed to determine optimal PCR cycles for linear amplification without saturating ATAC-Seq libraries. ATAC-Seq libraries were then quantified using Qubit (Invitrogen
identifiers.namespace: GEO
identifiers.id: GSM4671367