Table 2. Optimisation values for koala immune gene qPCRs developed in this study.
| Gene | Efficiency | Conc (nM) | Linear dynamic range (orders of magnitude) | LOQ (copies/μl) | Cq at LOQ | LOD (copies/μl) | r2 calibration curve | y intercept | Slope |
|---|---|---|---|---|---|---|---|---|---|
| CD4 | 1.02 | 500 | 9 | 4 | 36 | <1 | 0.99 | 43.09 | −3.27 |
| CD8β | 1.03 | 500 | 9 | 6 | 36 | <1 | 0.99 | 45.89 | −3.21 |
| IL-4 | 1.05 | 250 | 9 | 10 | 38 | 1 | 0.99 | 47.49 | −3.21 |
| IL-6 | 0.93 | 500 | 6 | 3358 | 36 | <1 | 0.99 | 43.54 | −3.49 |
| IL-10 | 1.06 | 250 | 9 | 0.7 | 36 | <1 | 0.97 | 48.75 | −3.17 |
| IFNγ | 0.92 | 300 | 6 | 37 | 33 | 3 | 0.98 | 39.97 | −3.52 |
| GAPDH | 0.97 | 500 | 10 | 3 | 38 | <1 | 0.99 | 42.26 | −3.40 |
| 28S | 0.97 | 250 | 9 | 2 | 35 | <1 | 0.98 | 38.89 | −3.39 |
Notes.
- Conc
- primer concentration (nM)
- LOQ
- limit of quantification
- LOD
- limit of detection