GSM8219561: TC5, SOS1i+MEKi, tumor3; Mus musculus; RNA-Seq
Tumors were harvested and processed on the gentleMACS Octo Dissociator. Digestion cocktail contained 800ug/ml Dispase II,100ug/ml DNase I and 400ug/ml Collagenase P. 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) according to the manufacturer's protocols & instructions. A total of 10,000 cells were targeted per library and processed in parallel. Any samples with a proportion >10% of cDNA 150bp-400bp, were subjected to an additional 0.6xSpri cleanup before proceeding with the remainder of library preparation. Libraries produced were checked for quality control using the Agilent Tapestation, quantified using the KAPA library Quantification Kit, diluted as appropriate, then sequenced using a NovaSeq6000 with paired-end sequencing and dual indexing as appropriate for the library preparation. Dual Index libraries were sequenced using cycling conditions 28, 10, 10, and 90 for Read 1, i7 index, i5 index and Read 2 respectively.