GSM9364366: Hashtag expression library from mouse sarcomas treated with vehicle or abemaciclib; GEM reaction 1; Mus musculus; OTHER
Tumors harvested when the average volume of the untreated group reached 1000 mm³, then were enzymatically and mechanically dissociated using the Mouse Tumor Dissociation Kit (130-096-730, Miltenyi Biotec) on the gentleMAC Octo Dissociator with Heaters (Miltenyi Biotec). Resulting cell suspensions were filtered through 70-µM strainers, and red blood cells were removed using ACK lysis buffer. Samples were hashtagged using antibody-conjugated oligo barcodes (TotalSeq-B hashtag reagents #1-8, Biolegend). Cells were stained with the viability dye Ghost Dye 710 (Cytek), and live cells were isolated by FACS. The resulting sample was concentrated to 1000 cells/uL and loaded into two reaction wells of the Next GEM 3' v3.1 scRNA-seq Chip G (10x Genomics) and captured on the Chromium X. Preparation of gene expression and hashtag libraries was executed according to manufacturer protocols for the Next GEM 3' v3.1 scRNA-seq kit (10x Genomics) and TotalSeq-B hashtags (Biolegend).