scRNA seq of mus musculus, 10w female bone marrow, vehicle, sorted CD3neg CD19neg
Single cell suspensions from each sample were prepared from spleen or bone marrow of SNS-ablated or control CT26-bearing wild-type BALB/c mice eighteen days after tumor challenge. Viable, CD3CD19 cells were sorted, washed, resuspended in PBS containing 0.04% BSA, counted, and captured for sequencing analysis. 12,000 cells were loaded into one lane of a 10X Chromium microfluidic chip. Single cell capture, barcoding and library preparation were performed using the 10X Chromium version 3 chemistry, according to the manufacturers protocol (#CG000183). cDNA and libraries were checked for quality on an Agilent 4200 TapeStation, quantified by KAPA qPCR, and sequenced on a single lane of a HiSeq4000 (Illumina) to an average depth of 50,000 reads per cell.