Transcription profiling of Herbaspirillum seropedicae SmR1 wild-type against its ntrC mutant under different nitrogen conditions
Transcription profiling of Herbaspirillum seropedicae SmR1 wild-type against its ntrC mutant under different nitrogen conditions
Cells were collected and stored in RNAlater stabilization solution (Thermo Fisher Scientific). Cells were cultivated in NFbHP-malate supplemented with 1 mM NH4Cl, at 30C and 250 rpm. When cells reached OD600 = 0.4, cells were collected and stored in RNAlater. Additionally, the remaining cells were incubated for a further 30 minutes after addition of 10 mM NH4Cl (Ammonium shock) or 10 mM KNO3 (Nitrate shock). After incubation, cells were collected and stored in RNAlater. RNA was extracted using Trizol (Thermo Fisher Scientific), and contaminant DNA was removed using TURBO DNase (Thermo Fisher Scientific). rRNA was removed using Ribo-Zero rRNA Removal kit (Gram-negative bacteria) (Illumina). cDNA libraries were constructed using Ion Total RNASeq kit v2 (Thermo Fisher Scientific).