ABSTRACT
The majority of the material that is described in this chapter will be devoted to demonstrating how to design and validate a new quantitative PCR (qPCR) assay. However, before reaching the point of designing a new assay, it is important to consider whether to take the plunge and design a custom assay or to order a premade assay from a commercial source. The rst step will be to go through that thought process, keeping the various potential goals of the research project in mind. Whether the decision is to purchase a predesigned assay or to make a new one, the validation steps described below will still need to be carried out. There are varying amounts of information about assay quality from the major sources of premade assays and even when it is available, you cannot assume that the assay will perform as expected in your hands, using your qPCR instrument with the reverse transcription (RT) and polymerase chain reaction (PCR) reagents that you have assembled.
