GSM4506452: WTinWT_Day21_rep2 [ATAC-Seq]; Mus musculus; ATAC-seq
To perform ATAC-seq, we FACS-sorted brain-infiltrating P14 cells (Tox +/+ and Tox -/-) from WT and MOG-GP mice at different time points after intracranial LCMV-GP infection. Each biological replicate (n = 2) represents 5x10E4 sorted P14 cells sorted in FACS buffer. Cells were spun down at 500 g for 20 min at 4 °C. Nuclei were isolated in 50 μl of lysing solution composed of 10 mM Tris-HCl, 10 mM NaCl, 3 mM MgCl2, and 0.1% IGEPAL CA-630. The nuclear pellet was resuspended into 50 μl of tagmentation buffer containing the Tn5 transposase (Illumina) and incubated at 37 °C for 30 min. The tagmented DNA was purified using MiniElute PCR Purification Kit (Qiagen). Libraries were preamplified for 5 cycles using indexed primer from Nextera kit (Illumina) and NEBNext High Fidelity PCR master mix (New England Biolabs). Preamplified libraries served as input to determine the number of remaining cycles needed for the second round of PCR amplification. ﻿Libraries were cleaned by Ampure Beads (Agencourt) at a 1.5× ratio.
GEO Accession
GSM4506452