GSM7832984: PP1-1_+Tet_B; Trypanosoma brucei; RNA-Seq
Total RNA was isolated from T. brucei cultures using TriPure, according to the manufacturer's instructions. 5x107 cells were used for each prep. For mRNA-seq, total RNA was isolated from T. brucei RNAi cultures grown in presence or absence of tetracycline for two days using TriPure. Six mRNA-seq libraries were constructed for each of the PP1 RNAi cell lines (triplicate samples for plus and minus tetracyclin) using Illumina TruSeq Stranded RNA LT Kit following the manufacturer's instructions with limited modifications. The starting quantity of total RNA was adjusted to 1.3 µg, and all volumes were reduced to a third of the described quantity. High throughput sequencing was performed on an Illumina Novaseq6000 instrument.