scATAC-seq of murine B cells post NP-OVA immunization

To determine the role of p52:ETS1 function in activated B cells. Spleen were harvested from WT and p52ki/ki mice (1 each) on day 9 post immunisation with NP-OVA and alum. B cells were isolated from total splenocytes and stained for FACs. Naive (CD19+IgD+) and activated B cells (CD19+IgD-) were sorted and mixed at 1:3 ratio. These sorted B cells (1 wt and 1 p52 ki/ki) were sent to immunogenomics facility at SIgN to be processed for single cell ATAC-seq on the 10x Genomics Chromium platform. Their protocol: Briefly, nuclei were prepared from sorted cells as per the 10x Genomics Demonstrated Protocol CG000169 followed by processing with the Chromium Single Cell ATAC Kit v2 following the manufacturer’s protocol, targeting the capture of up to 5000 nuclei. After quantification on the Agilent Bioanalyzer, ATAC-seq libraries were pooled for sequencing on the Illumina NovaSeq X, with the cycle configuration of Read 1N-151 cycles; i7 index-8 cycles, i5 index-16 cycles; Read 2N-151 cycles, targeting about 200 million reads-pairs/sample.