Illumina HiSeq 4000 sequencing; Exploiting evolutionary steering to induce collateral drug sensitivity in cancer
Exploiting evolutionary steering to induce collateral drug sensitivity in cancer
Single cells were prepared from POT, GEF1 and TRM4 cells. After centrifugation, single cells were washed with PBS and were re-suspended with a buffer (Ca++/Mg++ free PBS + 0.04% BSA) at 1,000 cells/µl. Viability was confirmed to be > 90% in all samples using acridine orange/propidium iodide dye with LUNA-FL Dual Fluorescence Cell Counter (Logos Biosystems, L20001). Single cell suspensions were loaded on a Chromium Single Cell 3' Chip (10X Genomics) and were run in the Chromium Controller to generate single-cell gel bead-in-emulsions using the 10X genomics 3' Chromium v2.0 platform as per manufacturer's instructions. Single-cell RNA-seq libraries were prepared according to the manufacturer's protocol and the library quality was confirmed with a Bioanalyzer High-Sensitivity DNA Kit (Agilent, 5067-4627) and a Qubit dsDNA HS Assay Kit (ThermoFisher, Q32851).
Experimental Factor: compound
trametinib
Experimental Factor: dose
100
Experimental Factor: time
63