Table 2. Checklist of tests to conduct when designing RT-qPCR studies.
| Checklist | Suggested Test |
|---|---|
| Correct reference genes | Test a panel of candidate reference genes using Normfinder |
|
Efficiency of primer
assays |
Conduct a calibration curve and use the slope of the graph to
calculate PCR efficiency with the following equation: 10 -1/slope-1 |
| Inhibition within samples | Add samples to a standard RT-qPCR reaction and look for
changes in the Cq values |
| RNA purity | Measure the ratio of absorbance at 260/280 nm |
| RNA integrity | Visualization of the 28S:18S ribosomal RNA ratio on a 1%
agarose gel |