GSM7664896: CD47 Knockout Replicate 1; Mus musculus; RNA-Seq
Spleens were removed from the mice and homogenized in HBSS and passed through a 70 µm mesh (Sigma, CLS431751) to remove debris. The cell suspension was treated with ACK lysis buffer for 4 minutes to lyse the RBC. The suspensions were centrifuged and washed twice with cold HBSS. Aliquots of single cell suspensions were stained using trypan blue and counted to assess viability. Samples were loaded according to the 10x 3' v3 User guide with a single capture lane per sample targeting a recovery of 6,000 cells per lane. Cell partitioning completed successfully with uniform emulsion consistency and the reverse transcription PCR was run overnight. All subsequent steps of library preparation and Quality Control were performed as described in the 10X Genomics 3' v3 Single Cell User Guide.