GSM9019564: RPM_Basal_Organoids_CMV_CellTagged_PostCre_GEX; Mus musculus; RNA-Seq
Organoids were subject to dissociation by TrypLE for 30 minutes into a single cell suspension. Allograft and primary tumors were subject to enzymatic/mechanical digestion into a single cell suspension. Select samples were multiplexed with 10x Genomics CellPlex technology. 10x Genomics gel beads and reverse transcription reagents were added to cell suspensions and loaded to a chromium single cell controller to form gel-bead-in emulsions (GEMs). From GEMs, cDNA was generated from barcoded mRNA and subsequent A tailing, end repair, adaptor ligation, and sample indexing was performed according to manufacturer. Libraries were assessed on Agilent D5000 ScreenTape on an Agilent Tech 2200 tapestation system and quantified by Kapa Biosystems Library Quantification Kit for Illumina Platforms. Libraries were sequenced on the Novaseq 6000, Novaseq X Plus, or NextSeq 1000 with paired end seq targeting > ~30k reads/cell for gene expression (GEX) libraries or ~5k reads/cell for CellPlexing/Cell multiplexing oligo (CMO) libraries. 10X Genomics Single-Cell 3' RNA Dual Index v3 and 10X Genomics 3' Feature Barcode Kit for CellPlexed samples