GSM7679951: mor28iCre-tdTom OMPtTA-tetOP2, tdTom+, ATAC, rep2; Mus musculus; ATAC-seq
ATAC-seq libraries were prepared from live sorted cells using the protocol developed by Buenrostro et al., 2015. Cells were pelleted (500 rcf, 5 min, 4°C) and then resuspended in lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630). Nuclei were immediately pelleted (1000 rcf, 10 min, 4°C). Pelleted nuclei were resuspended in transposition reaction mix prepared from Illumina Nextera reagents (for 50 uL: 22.5 uL water, 25 uL 2xTD buffer, 2.5 uL Tn5 Transposase). The volume of the Tn5 transposition reaction was scaled to the number of cells collected: 1 uL mix per 1000 cells. If fewer than 10,000 cells were collected by FACS, 10 uL scale reactions were performed. Transposed DNA was column purified using a Zymo DNA Clean & Concentrator-5 (Zymo). The transposed DNA was then amplified using barcoded primers and NEBNext High Fidelity 2x PCR Master Mix (NEB). Amplified libraries were purified using Ampure XP beads (Beckman Coulter) at a ratio of 1.6 ul of beads per 1 uL of library and eluted in 30 uL of elution buffer (10 mM Tris-HCl pH 8, 0.1 mM EDTA).