Figure 4.
Effect of the ref mRNA normalization on the 16S rRNA expression levels. Relative gene expression of the 16S rRNA normalized to the ref mRNA, using the delta Ct method, in 109 CFU/mL and 105 CFU/mL samples adjusted to the initial copies of ref mRNA used. For each bacterial concentration five technical replicates were spiked with standard concentrations of ref mRNA prior to RNA extraction and subsequent reverse transcription. For each condition two independent biological samples were used. Statistical significance within each samples’ set was determined by performing one-way ANOVA analyses followed by a Tukey’s multiple comparison test (****P < 0.0001).