RNA-seq for VBNC Corynebacterium diphtheriae

Transcriptional profiles for culturable, VBNC, VBNC + catalase (cells after 6 h adding catalase to VBNC at room temperature without light and agitation) and resuscitated (cells that were not subcultured following resuscitation) Corynebacterium diphtheriae were then evaluated as follows. C. diphtheriae from each state (culturable, VBNC, VBNC + catalase, and resuscitated) were suspended in 10 mL of PBS at an OD600 of 0.5. and collected by centrifugation at 3,000 ? g for 10 min and then used for total RNA extraction via a High Pure RNA Isolation kit. rRNA was depleted using the Bacteria Ribo-Zero kit and sequencing libraries were prepared using the TruSeq Stranded mRNA library Prep kit and sequenced on an Illumina NextSeq sequencer. cDNA quality and concentration were evaluated using the Agilent High Sensitivity DNA kit and the Agilent 2100 Bioanalyzer. Paired-end reads with a length of 2 ? 75 bases were used to generate the whole transcriptome libraries.