GSM8772543: 20220531_hpap_2_Pool_Run1_Well2_ADT, ADT derived library.; Homo sapiens; OTHER
Pooled samples were placed through the 3' scRNAseq v3.1 pipeline (10X Genomics) to generate single cell partitions sharing cDNA, antibody derived tag, and hashing antibody barcodes. Each well of a 10X chip was superloaded with ~1.75X manufacturer's recommended maximum cell count. cDNA extract protocol followed manufacturer's instructions. To extract antibody derived tag and hashing tag sequences, the supernatant from cDNA Cleanup - SPRIselect (Step 2.3d in Protocol CG000315 Rev C, or equivalent step) was saved for further processing following the standard CITEseq protocol (https://cite-seq.com/protocols/). Dual indexed cDNA libraries were generated following manufacturer's instructions (10X Genomics Protocol CG000315 Rev C, or equivalent for high-throughput protocols). Dual indexed antibody derived tag and hashing tag libraries were generated following standard CITEseq protocol (https://cite-seq.com/protocols/) using primers recommended from Biolegend. Antibody derived tag and hashing tag sequences were enriched in the initial cDNA amplification step of the 3' scRNAseq v3.1 protocol via addition of additive primers (https://cite-seq.com/protocols/).