GSM3380364: ATAC_MV_1; Mus musculus; ATAC-seq
Protocol for transposition reaction and PCR based on (Buenrostro et al., 2013). 50,000 nuclei per condition where pelleted and resuspend in 1XTD Buffer (Illumina) with 2.5ul Tn5 transposase (Illumina) in 50µl volume, at 37°C 300rpm for 30min. DNA was purified by MiniElute PCR purification (Qiagen), before test amplification to calculate library amplification cycle number with NEBNext® Ultra™ II Q5 Master Mix (NEB), Sybre green and customized Nextera PCR Primers (Buenrostro et al., 2013) using a LightCycler480 (Roche). ATAC-seq libraries where uniquely indexed with customized Nextera PCR Primers and amplified with 11 cycles of amplification before QIAquick PCR purification (Qiagen), and Ampure XP Bead (Beckman Coulter) size selection, and sequenced on an Illumina HiSeq 4000 75bp PE sequencing.
GEO Accession
GSM3380364