RNAseq of isogenic RPE1 cells
Total cellular RNA will be extracted and the quality will be assessed using the Bioanalyzer 2100. For each sample, total RNA will be depleted of ribosomal RNA and the RNAseq libraries will be prepared with the Illumina TruSeq Stranded Total RNA kit following the manufacturers protocol. Briefly, after the fragmentation of RNA using divalent cations at elevated temperature, cDNA will be synthesized, end-repaired and 3-end-adenylated. Following adapter ligation, libraries will be amplified by PCR. Amplified libraries will be checked on a bioanalyzer 2100 and quantified with picogreen reagent. Libraries with distinct TruSeq adapter UDIndexes will be multiplexed and after cluster generation on FlowCell will be sequenced for 50 bases in the paired-end mode with 35 Million reads in coverage on a Novaseq 6000 sequencer