Illumina NovaSeq 6000 paired end sequencing; scRNAseq of BRAF-mutant Colo205 cells control or treated for 24h or 72h with vemurafenib
scRNAseq of BRAF-mutant Colo205 cells control or treated for 24h or 72h with vemurafenib
Samples were grown in RPMI1640 with 10% FBS and treated as indicted before harvest by trypsin-based detachment for scRNA-seq. Cells were subjected to gel-in-emulsion capturing using the 10x Genomics protocols as single cells with beads containing reverse transcription primers and reagents. cDNA was constructed using primers containing 10x Genomics GEM beads containing Illumina R1 sequence, 10x Barcode, a UMI and poly-dT sequences. Barcoded cDNA was PCR amplified, purified and R2 sequencing primers were added before sequencing.