Study: Whole CD45+ splenocytes from B6 mice, 10X (Broad) 5758 cells
Whole CD45+ splenocytes from B6 mice, 10X (Broad)
As a first pass to develop common Standard Operating Protocols to produce compatible single-cell RNAseq across the consortium, Immgen labs (Wash U. St Louis, Broad, HMS) generated parallel datasets from spleens of un-perturbed C57Bl/6 mice.
Per ImmGen SOP, mice were 5 w.o. C57Bl/6J (B6) mice bred at JAX, shipped one week prior and maintained in SPF conditions. Spleens were mechanically dissociated, and CD45+ cells were purified by flow cytometry (Aria) immediately prior to encapsulation on a 10X genomics instrument. Libraries were constructed with the Chromium Single Cell 3’ Library Kit. The libraries use a KAPA Library Quantification Kit for Illumina platforms for the post construction quantification.
After QC and selection of cells passing depth criteria (>250 genes/cell), we obtained valid results for 3025 and 2733 cells from female and male B6 donors, respectively.