RNAseq of splenic germinal center B cells from BM Only control transplanted mice
Day 49 post-BMT spleens were homogenized, RBC lysed, stained for FACS sorting with fixable-viability dye, anti-CD19, anti-GL7, and anti-FAS (same products as above) and cells collected at 100,000 targeted number directly into an equivalent volume of Qiagen buffer RLT (Qiagen 74134) + 1% 2-ME (Sigma M6250-100ML). Samples were frozen on dry ice until final processing at the University of Minnesota Genomics Center. RNA was collected and isolates quantified by fluorometric RiboGreen assay, integrity determined using capillary electrophoresis and samples converted to Takara sequencing libraries using Takara Bios SMARTer Stranded Total RNA-Seq  Pico Mammalian Kit v2 (Cat. # 634414).