GSM8995449: R255X-WT_CO1, R255X-MUT_CO1, R255X-WT_TA1, R255X-MUT_TA1; Homo sapiens; RNA-Seq
Multiplexing was performed with antibody-based cell hashing. Library preparation was performed according to the manufacter's instructions (GEM-X Single Cell 3' Kit v.4 protocol, 10x Genomics). Antibody-based cell hashing was performed using 0.5 µg of TotalSeqTM-B anti-human antibodies against β2-microglobulin and CD298 (BioLegend: 394631, 394633, 394635, and 394637), according to the manufacturer's instructions. To generate single-cell GEMs (Gel Beads in Emulsion), 40,000 live cells of the multiplexed pool were loaded per channel (to give an estimated recovery of 10,000 cells per channel) onto a Chromium GEM-X Single Cell Chip (10X Genomics, PN-1000690). The gene expression library was prepared with the GEM-X Single Cell 3' Kit v.4 (10X Genomics, PN-1000691), and the hashtag-oligo library was prepared with the GEM-X Single Cell 3' Feature Barcode Kit (10X Genomics, PN-1000702). ScRNA-seq libraries were then pooled and pair-end sequenced in a NovaSeq X flowcell (Illumina) with a targeted sequencing depth per cell of 50,000 gene expression reads and 5,000 surface barcode reads.