GSM9285335: PP_MBC_Tneg_611; Mus musculus; ATAC-seq
Tissue were harvested and cells of each immune population were sorted into tdTomato positive and negative fractions representing early life origin B cells or non-early life origin B cells, respectively. Cells were centrifuged for 10 minutes at 500xg, and tagmentation was performed by resuspending cells in 25 ul of Tagmentation mix (1X TD buffer (10 mM TAPS-NaOH pH 8.1, 5 mM MgCl2, 10% DMF), 2.5 μl Tn5 transposase, 0.1% Tween 20 and 0.02% Digitonin) and then incubated for 1 h at 37°C. DNA was purified and amplified into a sequencing library with KAPA HiFi Polymerase and the Nextera Indexing Kit Primers (Illumina, Inc).