PL-KO (CRX856670)
Cellular suspensions were loaded onto a 10X Genomics GemCode Single-cell instrument to generate single-cell Gel Bead-In-EMulsion (GEMs).  Libraries were generated and sequencedfrom the cDNAs with Chromium Next GEM Single Cell 3'Reagent Kits v3.1.  Silane magnetic beads were used to remove leftover reagents and primers, and full-length, barcoded. cDNAs were amplified by PCR for library construction.  The libraries were constructed by adding specific primers during GEM incubation and library preparation steps, including End Repair, A-tailing, Adaptor Ligation, and PCR. To investigate the involvement of FAM3D in neutrophil heterogeneity during infection, single-cell RNA sequencing (scRNA-seq) was performed on peripheral blood (PB) and peritoneal lavage fluid (PL) samples collected from both WT and Fam3d-/- mice following E. coli-induced peritonitis. Our samples were collected 4 hours post-infection.