scRNA-seq of human B cells stimulated in vitro with CpGC
scRNA-seq of human B cells stimulated in vitro with CpGC
Blood into Lithium Heparin, PBMC prep by Ficoll Paque, B Cell isolation by EasySep B cell Negative Selection Kit. 1uM CpGC stimulation for 72h.   Blood into Lithium Heparin tubes, PBMC prep by Ficoll Paque. Cells frozen in 10% DMSO RPMI complete medium and defrosted when required. CD19 Miltenyi isolation of B cells.   10X genomics 5' V1, in drop lysis and oligo(d)T reverse transcription with template switch oligos. as per manufacturer's instructions 10X genomics 5' V1, single index as manufacturer's instructions IL-10 was specifically amplified cDNA for the CpGC samples using primers forward 5'GCCTAACATGCTTCGAGATC3' and reverse 5'TGATGTCTGGGTCTTGGTTC3'.  Stardard i7 indexing was then used to produce the library.