Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: 1 ml of Trizol was added to cells and cells were frozen at -80°C for at least 1h. Subsequently cells were thawed at 37°C. RNA release was performed using silica beads and using the Precellys 24- Dual cell homogenizer (Peq-Lab) twice for 20 s at 5000 rpm. Samples were incubated for 15 min at room temperature. Subsequently, 100 µl 1-bromo-3-chloropropane was added to the samples and centrifuged for 15 min at 8000 rcf at 4°C to separate aqueous and organic phase. The aqueous phase was used to isolate total RNA with the RNeasy mini kit (Qiagen) following manufacturer instructions. RNA libraries were prepared using the RNAeasy mini kit (Qiagen)