Table 3. 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% non-denaturing agarose gel