GSM9726201: TC4, FAP TcE, tumor1; Mus musculus; RNA-Seq
Tumors were harvested and processed on the gentleMACS Octo Dissociator. Digestion cocktail contained 800ug/mL Dispase II (GIBCO), 400ug/mL Collagenase P (Roche/Sigma), and 100ug/mL DNase I (Roche/Sigma). Cell viability was assessed with Zombie Yellow Fixable viability dye. Single cell sequencing library preparation was performed using the 10x Genomics Chromium Controller and Chromium Next GEM Single Cell 3' Kit v3.1 Chemistry Library, Gel Bead, and Chip Kits (10x Genomics). 10,000 cells were targeted per library and processed in parallel with quality control performed using Agilent Tapestation. Library quantification used the KAPA library Quantification Kit and was sequenced by NovaSeq6000 with paired-end sequencing and dual indexing as appropriate for the library preparation. Cycling conditions for single index libraries were 28, 8, and 91 cycles for Read 1, i7 index and Read 2 respectively. Dual Index libraries were sequenced using cycling conditions 29, 10, 10, and 91 for Read 1, i7 index, i5 index and Read 2 respectively.