GSM8107979: cDNA, 10X Lane1, Sequencing Run 2; Homo sapiens; RNA-Seq
Each transduced cell line was split into four batches of 500,000 cells, resulting in a total of 8 different hashes. After incubation and washes, all 8 hashed batches were pooled together and run in two reaction lanes of the 10X Chromium Next GEM Single Cell 5' Reagent Kit v2 (single indexing, PN-1000265 and PN-1000190). The manufacturer's protocol was followed with modifications stipulated in the ECCITE-seq protocol 11. For each GEM reaction, 42,000 cells from the hash pool were used to obtain approximately 21,000 total cells, including "multiplets" (multiple cells per droplet counts) Gene expression (cDNA), hashtags (HTOs), and guide RNA (Guide-derived oligos, GDOs) libraries were constructed following the 10x Genomics and ECCITE-seq protocols (https://cite-seq.com/eccite-seq/) with minimal modifications. Specifically, the antibody pool protein tag library steps were ignored, and a custom-designed probe library was used to enrich the cDNA for the genes of interest in the 10X Targeted Gene Expression protocol (10X PN-1000248)