Table 1.

RT-qPCR data and fold difference calculations. Two independent RT-qPCR runs were performed, each with two RNA extractions that each tested for 16s and mecA expression levels in triplicate. Using the comparative quantification standard curve method, the fold difference in bacterial gene expression was given by the ratio of the efficiency of the target gene raised to the ΔCt of the target to the efficiency of the normalizer gene raised to the ΔCt of the normalizer. Efficiencies were calculated from the slopes of the standard curves. Calculations showed an average fold difference of 0.230, meaning that the S43 CRISPR-dCas9 treated MRSA had a significant decrease of 77% in mecA gene expression.

SampleGeneCt NormalizerCt TargetCt SampleΔCt NormalizerΔCt TargetSlope