Definitive qPCR

An exhaustive guide to assay design for quantitative real-time PCR. The book describes the basic concepts important for amplicon selection and primer and probe design. There are step-by-step examples for designing probe-based and SYBR Green assays targeting mRNA and fungal pathogens using several popular design programs. These are then exposed to extensive in silico analysis to identify the optimum amplicon/primer/probe combination. There is a detailed trouble shooting guide, a listing of instruments, reagents and additional information available on the internet, all with hyperlinks. In addition, there are three Keynote presentations summarising the main concepts of standard assay design, multiplex assay design and explaining the rationale behind MIQE, the guidelines for qPCR publication transparency.