GSM8896066: CaCo-2/HT29-MTX 9:1 co-culture_subletal toxin concentration_2h incubation, biol rep 3; Homo sapiens; RNA-Seq
Total RNA preparation and on-column DNase digestion were performed using the „Nucleospin RNA II" kit (Machery Nagel) according to the instructions of the manufacturer. RNA quality was tested via spectrophotometer and agarose gel electrophoresis. RNA purity was confirmed via PCR for ITS1 and 18 S rRNA as control. Whole transcriptome sequencing via rRNA depletion: Ribosomal RNA depletion -> RNA fragmentation and random priming -> First and second strand cDNA synthesis -> End repair, 5' phosphorylation and dA-tailing -> Adaptor ligation, PCR enrichment and sequencing