Illumina HiSeq 1500 paired end sequencing; Differential RNA-Seq of Corynebacterium glutamicum wildtype, deletion mutant sigE and deletion mutant cseE reveals overlapping SigH and SigE sigma factor regulons.
Differential RNA-Seq of Corynebacterium glutamicum wildtype, deletion mutant sigE and deletion mutant cseE reveals overlapping SigH and SigE sigma factor regulons.
The cells were harvested in the exponential growth phase and frozen in liquid nitrogen. Total RNA was isolated from 3 biological replicates of C. glutamicum cells by a Quick-RNA Miniprep Plus kit (Zymo Research). The samples were treated with DNase (Roche Diagnostics) and RNA was purified with an RNA Clean&Concentrator-5 kit (Zymo Research).  Ribosomal rRNA was removed with a Ribo-Zero rRNA Removal Kit for bacteria (Illumina). The purity of RNA was then tested with an Agilent RNA Pico 6000 kit and an Agilent 2100 Bioanalyzer (Agilent Technologies). TruSeq Stranded mRNA Sample Preparation guide (Illumina) was then used to construct the cDNA library.