GSM7989942: Islet_168 (SAMN29494287), HTO; Homo sapiens; RNA-Seq
Islets were treated with Accutase for 8-10 min at 37°C to generate a single cell suspension. Cells were then washed and suspended in Staining buffer (PBA, 2%BSA, 0.01%TweenS). Islets were incubated with Fc Blocking reagent (FcX, BioLegend) 10 minutes at 4 ̊C, before adding 0.5ug of a unique Cell Hashing antibody (TotalseqTM-A0251 to A0257 anti-human hashtag antibody, Biolegend) for 20 minutes at 4 ̊C, and then washed 2 times with Staining buffer, and once with PBS+0.04%BSA. Cell viability was assessed on a Countess II automated cell counter (ThermoFisher), and up to 30,000 cells (~5,000 cells from each hash-tagged (HTO) sample) were loaded onto one lane of a 10X Chromium Controller. One single cell suspension was loaded twice i.e. across 2 lanes on the 10x chip, and 2 gene expression and 2 HTO libraries were generated in total, which were processed into a single set of gene expression and HTO outputs. Single cell capture, barcoding and library preparation were performed using the 10X Chromium platform V3 chemistry and according to the manufacturer's protocol (#GC000103).