Fig. 6

Experimental determination of transcriptional orientation by two-step RT-PCR and verification of biofuel-responsive sRNA. a Four biofuel-response sRNA orientations by two-step RT-PCR. “+” denotes the orientation of sRNA on the positive genome strand, “−” denotes the orientation of sRNA on the negative genome strand. The bold and underlined names indicate the determined orientation of an sRNA. b Ethanol- and butanol-responsive expression of Nc117 validation using two-step RT-PCR. The upper part indicates Nc117 validation, the control background primer-free or RNA-free for Nc117 were conducted with no primers or reverse transcriptase added during first step reverse transcription. The lower part shows 16S rRNA indicates an internal control, all samples with random primers during reverse transcription, except for lane 8 RNA-free without reverse transcriptase. All samples were collected in the logarithmic growth phase