GSM9203999: Hashtags; Homo sapiens; RNA-Seq
CAR T cells were oligo-tagged using TotalSeq A anti-human antibodies (BioLegend): Hashtag 5- AAGTATCGTTTCGCA (CAT#394609), Hashtag 6- GGTTGCCAGATGTCA (CAT#394611), Hashtag 7- TGTCTTTCCTGCCAG (CAT#394613), Hashtag 8- CTCCTCTGCAATTAC (CAT#394615) and combined into a single suspension. Single Cell RNA-seq and Cell Surface Protein libraries were constructed using the Chromium Single Cell 3′ v3.1 Reagent Kit (10x Genomics, PN-1000268) according to the manufacturer's protocol (manual part number CG000204 Rev D). Reverse transcription and library preparation were performed on a C1000 Touch Thermal cycler with 96-Deep Well Reaction Module (Bio-Rad).  cDNA was amplified 12 cycles and evaluated on a Agilent BioAnalyzer 2100 using a High Sensitivity DNA Kit (Agilent Technologies).  Final Gene Expression and Cell Surface Protein libraries were amplified 12 and 10 cycles respectively and visualized on an Agilent TapeStation 4200 using High Sensitivity D1000 ScreenTape (Agilent Technologies).  Individual libraries were diluted to 2nM and pooled for sequencing.