GSM6327100: Treg, Il10 negative, mRNA-derived cDNA [E2_Il10neg]; Mus musculus; OTHER
Cells were sorted into 0.04% BSA/PBS, pelleted, and then resuspended at 1000-1500 cells/ul Library was prepared using Chromium Single Cell 3ʹ v3 protocol (10x Genomics). Cellular mRNA and antibody-derived oligos were reverse-transcribed and indexed with a shared cellular barcode. Indexed cDNAs were then pooled and amplified by PCR according to the Chromium Single Cell 3' v3 protocol (10x Genomics) and with specific primers amplifying the antibody-derived tags. SPRI bead size selection was performed in order to separate both the mRNA-derived cDNA (>300bp) and the antibody-derived tagged cDNAs (180bp). For the mRNA derived cDNA library preparation, we further proceeded according to the manufacturer's instructions for Single Cell 3' v2 protocol (10x Genomics). For antibody-derived tagged library, we followed the protocol provided by BioLegend (HTO, TotalSeq-A)