title: GSM6674215: BM23-LN2-GEX Lymph node mononuclear cells, mRNA library, replicate 2; Macaca mulatta; OTHER
library_construction_protocol: Single cell suspensions were prepared from whole femur bone marrow, lymph nodes, and peripheral blood samples obtained from each rhesus macaque. Gradient density centrifugation was used to fractionate each single cell suspension and mononuclear cells were collected. Mononuclear cells were treated with ACK lysis buffer to remove red blood cells. Purified mononuclear cells were enumerated and 1*10^6 cells from each sample were stained with a panel of 124 oligo-conjugated antibodies targeting cell surface proteins or isotype controls (TotalSeqA; Biolegend). Following antibody staining, cells were loaded into a 10x Genomics Chromium Instrument. Two biological replicate samples per tissue per animal were generated targeting a cell recovery of 10,000 cells per sample. Libraries were generated using 10x Genomics 3' scRNAseq v3.1 kit with feature barcoding per manufactuer's recommendations. ADT libraries were contructed according to manufacturer's recommendations using the TotalSeqA Reagent Kit v3 3.1 (Biolegend). Final libraries were pooled at a ratio of 80:20 RNA:ADT prior to sequencing. 100,000 reader per cell were targeted for RNA libraries and 25,000 reads per cell were targeted for ADT libraries. OTHER: CITE-seq (Cellular Indexing of Transcriptomes and Epitopes by sequencing)