RNA-seq of BRAF-V600E_mutated and BRAF-V600E_corrected colon cancer organoids
RNA-seq of BRAF-V600E_mutated and BRAF-V600E_corrected colon cancer organoids
Organoids were collected from Matrigel by enzymatic digestion using 1 mg/ml dispase for 10 minutes at 37°C and washed 2 times in 1xPBS. Organoids were lysed in RLT-buffer (Qiagen) supplemented with 1% β-mercaptoethanol and were stored at -80C. Organoids were grown in the presence of EGF-supplemented medium for 5 days Total RNA was extracted using the QIAGEN RNeasy Plus Micro kit as per manufacturer's instructions. DNase (Qiagen) treatment was included to avoid possible genomic DNA contamination. RNA was eluted in 30 μl of RNAse free water. Libraries were prepared using Truseq RNA stranded polyA (Illumina) according to the manufacturer's directions.