Project description:Transcriptional profiling of Corynebacterium glutamicum cells comparing wild-type cells with cg0196 deletion mutant cells by site-specific gene deletion using the non-replicable integration vector. cg0196 is gene conding transcriptional regulator related carbon metabolism.
Project description:Total RNA was isolated from 3 biological replicates of C. glutamicum wildtype, deletion mutant sigE and deletion mutant cseE 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. The constructed cDNA library was then sequenced with Illumina HiSeq 1500 using a read length of 74 bases.
