GSM9181234: scRNA-seq, P04, before RTX treatment; Homo sapiens; RNA-Seq
Frozen PBMC samples were thawed rapidly in trypsin-stopping solution (D-MEM Low Glucose, 10% FBS, and 1% penicillin/streptomycin) at 37°C, centrifuged at 400g for 10 minutes at 4°C and the supernatant was discarded. To release cell aggregation, 25 μL of DNase Booster Buffer and 10 μL of DNase I stock solution (#79254, QIAGEN) were added to the pellet and incubated at room temperature for 15 minutes. The samples were washed with trypsin-stopping solution and diluted with PBS. To isolate CD3+ T cells, PBMCs collected at both pre and post were stained and sorted at 4°C using pooled monoclonal antibodies and sorted using FACSAria Fusion flow cytometer (BD Biosciences). Single-cell 5' and TCR libraries were prepared following the protocol provided by the 10x Genomics Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index).