CITE-Seq of homo sapiens: adult female peripheral blood and ovarian follicular fluid
Peripheral blood mononuclear cells (PBMC) and follicular fluid mononuclear cells (FFMC) were separated from plasma and follicular fluid respectively by density gradient centrifugation and washed with RPMI + 10% fetal bovine serum. ACK lysis buffer was added to FFMCs to remove red blood cell contamination. Samples were enriched for live cells and single-cell suspensions were prepared. FFMCs and PBMCs were separately incubated with multiplexing antibodies and then pooled. All samples were then incubated with selected TotalSeq B Feature Barcode antibodies. Single cells were separated and cDNA libraries prepared via PCR. Feature Barcode/CITE-Seq antibody cDNA was separated.