Fig. 2

Illustration of the flow of the amplification and melting curve analysis. At different steps in data processing and analyses (rectangular boxes) the program can detect deviating reactions and provide warnings or errors (tabbed boxes). The flow chart gives a suggestion on how the user can deal with those warnings or on how to improve the assay to avoid these errors in the future (rounded boxes). Note that these recommendations are not exhaustive; basic knowledge on qPCR assay design and data analysis, as well as papers on these subjects, should guide the user in the appropriate direction. PC: positive control; NC: negative control; Unk: unknown sample;