title: GSM8170748: PB, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: The 6-9 week old C57BL/6J mice were endotracheally innoculated Klebsiella pneumoniae (ATCC 43816) to induce sepsis. PBMCs and BMMCs were separated with the 1.084 g/mL Ficoll medium. In brief, EDTA-anticoagulated whole blood and BM single-cell suspension diluted with an equal volume phosphate-buffered saline (PBS), spread flat on the surface of Ficoll layer and centrifugated at 350 g for 30 min. Next, the middle PBMC and BMMC layer was retained and erythrocytes in it was lysed using lysis solution at room temperature for 5min. After lysis terminating and cell washing with precooled PBS adding 2% fetal bovine serum, cell counting and viability were assessed using trypan blue by microscopy. Finally, once ensuring that the cell viability exceeded 88%, isolated PBMCs and PBMCs were frozen in liquid nitrogen until use. Prepared single-cell suspension is partitioned into GEMs (Gel Beads in Emulsions) in the automated Chromium Controller, and then mRNAs are reverse transcribed into cDNAs. Library was performed according to the manufacture's instruction (single cell 3' v2 protocol, 10× Genomics).