There are currently two published articles that discuss the background and operation of the software:

Typically, samples obtained from studied organisms are processed in several preparative stages before being analysed using qPCR:

1)   subjects are randomly selected from the population

2)   mRNA samples withdrawn from each subject

3)   RT products obtained from each sample

4)   qPCR analyses performed on each RT

This hierarchy is susceptible to the introduction of processing errors at each level that will propagate down through successive levels and be reflected in the observed qPCR result.  The processing errors present in the final result serve only to confound the interesting biological variation, such as the effect of a treatment or the variation between subjects.  It is therefore the prerogative of the experimenter to design the assay in such a way as to minimise the scope for the introduction of this confounding variation.

To create the optimal experimental design, knowledge of the sources of error throughout the sample processing stages is essential.

In a given dataset, powerNest will assess the variance at each of these levels and use this information to produce an optimal design.

Sampling procedure

qPCR sampling

further reading