Table 1 List of candidate reference genes (RGs). The amplicon size, PCR efficiency percentage, minimum and maximum threshold cycle (CT) values observed across all tested growth conditions, the standard deviation of CT values across all tested growth conditions (determined by BestKeeper), and the CT value of the no template control (NTC) are listed for each RG. See Supplementary Figure 1 for standard curves used to calculate PCR efficiency. See Supplementary Figure 2 for melting curve analysis. See Supplementary Table 1 for oligonucleotide sequences and melting and annealing temperatures.
From: Selection of stable reference genes for RT-qPCR in Rhodococcus opacus PD630
Reference gene (RG) | Gene number | Gene annotation | Amplicon size (bp) | PCR efficiency | Min CT | Max CT | CT Std Dev | NTC CT |
|---|---|---|---|---|---|---|---|---|
RG1 | PD630_ RS22865 | Pup-protein ligase | 157 | 94% | 23.44 | 25.23 | 0.40 | N.d. |
RG2 | PD630_ RS20570 | Hypothetical protein | 87 | 94% | 25.86 | 28.20 | 0.67 | N.d. |
RG3 | PD630_ RS03840 | 23S rRNA | 87 | 95% | 8.74 | 10.16 | 0.33 | N.d. |
RG4 | PD630_ RS15810 | Polyribonucleotide nucleotidyltransferase | 115 | 101% | 22.93 | 27.41 | 1.37 | N.d. |
RG5 | PD630_ RS25785 | L,D-transpeptidase Mb0493 | 90 | 101% | 24.56 | 25.96 | 0.37 | N.d. |
RG6 | PD630_ RS13910 | NAD(P)H dehydrogenase | 82 | 100% | 25.47 | 29.01 | 1.13 | N.d. |
RG7 | PD630_ RS25530 | ATP-dependent Clp protease ATP-binding subunit ClpX | 94 | 92% | 24.93 | 25.77 | 0.21 | N.d. |
RG8 | PD630_ RS01395 | 16S rRNA | 101 | 92% | 10.48 | 11.74 | 0.29 | N.d. |
RG9 | PD630_ RS37755 | rRNA small subunit methyltransferase G | 102 | 92% | 28.32 | 30.33 | 0.46 | N.d. |
RG10 | PD630_ RS27310 | DNA polymerase IV | 71 | 94% | 28.34 | 30.61 | 0.61 | N.d. |
