Study: Whole CD45+ splenocytes from B6 mice, 10X (HMS) 10651 cells

Whole CD45+ splenocytes from B6 mice, 10X (HMS)

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 10651 cells from a B6 female donor.