GSM8169921: 2913_RK_Rep1_MinusDox; Trypanosoma brucei; RNA-Seq
For RNA isolation, doxycycline-induced and uninduced cells were harvested, Trizol reagent (Invitrogen) was added and this suspension was frozen at −80℃. Total RNA was isolated using Trizol reagent and phenol–chloroform extraction followed by DNase-I treatment (DNA-free DNase kit; Ambion). The RNA was further purified using phenol–chloroform extraction and the purity and quality of the RNA were measured using Nanodrop 1000, and the 260/280 ratio was ~2. RNAseq was done using the PolyA mRNA magentic isolation beads (E7490) and the NEBNext Ultra II directional RNA kit (E7760) from NEB.  Samples were sequenced on a NextSeq2000, PE100,  P3 flow cell.