GSM8219520: PFF-AZD_GEX; Mus musculus; RNA-Seq
Mononuclear cells were isolated for sequencing as described above with 3-5 ventral midbrains pooled per sample and sorted for CD45+ live cells on a BD FACsAria. Sorted CD45+ cells were resuspended in DPBS supplemented with 0.04% w/v BSA. Single Cell 3′ Reagent Kit V3.1 was used in conjunction with TotalSeqB antibodies to generate 3' gene expression library and HTO library. Single cells and beads containing barcoded DNA oligos were captured into oil droplets to generate the gel bead-in-emulsion (GEM) by 10x Chromium controller. The poly-A RNA from the cell lysate in GEM was reverse-transcripted to cDNA. Libraries were constructed according to the manufacturer's instructions