RNA-Seq of wild type replicate 1
Briefly, 1000ng of RNA sample was used for ribodepletion by using NEBNext rRNA depletionkit Bacteria (Cat. no. NEB E7850X), which includes Probe hybridization of RNA, RNase Hdigestion, DNase 1 digestion according to the manufacturers protocol. The NEBNext UltraII Directional RNA Library prep kit for Illumina (Cat. no. E7760L), was used to construct double-stranded cDNA libraries from the rRNA depleted RNA, using RNA fragmentation, first strandcDNA synthesis with random primers, second strand cDNA synthesis, end repair of doublestranded cDNA, adapter ligation, removal of excess adapter using sample purification beads,PCR enrichment of adapter ligated DNA and clean-up of PCR products as instructed by themanufacturer. The cleaned libraries were quantitated on Qubitflurometer and appropriatedilutions loaded on a D1000 screen tape to determine the size range of the fragments and theaverage library size.