GSM5235545: Multiplexed sample; Homo sapiens; RNA-Seq
For assessment of sub-clonal transcriptional identity parental and daughter TP0 and TP1 clones were expanded, following which 0.5x106 cells from each population of interest were washed and incubated with 5ul of Trustain FcX for 10 minutes on ice (Biolegend, #422302). To minimize batch effect, all 12 samples were prepared in parallel and subjected to cell hashing. Each population was incubated with a unique combination of 2 TotalSeq-C antibodies for 3 minutes on ice (Biolegend). Cells were washed twice in cold PBS-0.04% BSA and normalized before being mixed in equimolar amounts and resuspended to 1x106/ml. 20,000 cells were loaded onto a single 10X channel using the Single Cell 5' reagent kit. Sequencing libraries were subsequently constructed using reagents from a Chromium Single Cell 5' reagent kit according to the manufacturer's instructions. Sequencing was performed on an Illumina NovaSeq platform (paired-end) according to the manufacturer's instructions (Illumina).