Dissection of memory response B cell populations by single cell RNA-seq

SWHEL B cells were adoptively transferred and recipient mice were immunised with hen egg lysozyme (HEL) to establish anti-HEL memory B cells. Immune mice were then re-immunised with HEL and single donor-derived antigen-specific B cells sorted for RNA sequencing on day 5 of the secondary antibody response. Single-cell resolution transcriptional profiles of 144 memory response (Day 5 post-recall) SWHEL-Tomato B cells from three mice. Single-cell samples were isolated by FACS and processed in 9 batches. Samples were split into two pools of multiplexed samples. Each pool was run on a single Illumina HiSeq 2500 lane, averaging 3 million reads per cell.