RNA-seq to investigate how small RNA ncS27 in Burkholderia cenocepacia regulates uptake and metabolism of carbon compounds

Burkholderia cenocepacia small RNA ncS27 was identified by transcriptome profiling. To investigate the function of this small RNA, ncS27 was overexpressed and silenced in trans and the effect of this on the transcriptome was studied. To overexpress ncS27 it was cloned into a vector with a rhamnose inducible promoter. To silence ncS27 it was cloned into the same vector, but in reverse orientation. The vectors were then transformed into B. cenocepacia J2315, creating two mutants. The third mutant in this experiment is a vector control mutant. All three mutants were cultivated in Lennox broth to OD 0.5, then the inducer rhamnose was added. After 30 min of incubation in presence of the inducer the cells were harvested, RNA was extracted and RNA-sequencing performed.