Structural changes in the transcriptional response to N deprivation promote nitrogen cost minimization in Prochlorococcus
Total RNA was treated with Ribo-Zero rRNA Removal Kit for bacteria (Epicentre) to remove rRNA. The samples were next treated with 5' RNA polyphosphatase (5RPP; Epicentre) in order to convert 5' triphosphate (5'PPP) structures into 5' monophosphate ends. 5’ Illumina TruSeq adapters were ligated to the monophosphate groups of the transcripts and cDNA was synthesized. The tagged 5' cDNA fragments were then specifically amplified with PCR and sequenced using 5’ RACE tagRNA-seq on a NextSeq 500 system with 75 bp read lengths (vertis Biotechnologie AG)