GSM8507253: BCG neonatal 1; Mus musculus; RNA-Seq
Splenocytes from naïve, BCG-vaccinated, septic (18 hours post-CS) and BCG plus sepsis (18 hours post-CS) neonatal mice, were obtained after mechanical dissociation using 70 mm strainers.  After centrifugation at 300 x g for 10 min, RBC were lysed with ACK lysing buffer (Thermo Fisher Scientific, Waltham, MA) for five minutes at room temperature.  Total splenocytes were washed in PBS supplemented with 2% FBS, quantified (Cellometer Auto 2000 Cell Viability Counter; Nexcelom Bioscience, Lawrence, MA) and diluted to 106 cells/ml before single-cell library preparation. Libraries were prepared with Chromium Next GEM Single Cell 5' v2 kit targeting 10,000 cells/lane on the 10x Chromium microfluidics device (10x Genomics, Pleasanton, CA).  Libraries were sequenced at a minimum depth of 20,000 read pairs per cell on an Illumina Novaseq X plus Platform (Illumina, San Diego, CA).