Practical qPCR: Assay Design

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.

Contents: Introduction Why assay design is important Why design your own assay? Tools for assay design Amplicons Amplicon selection – mRNA Amplicon selection – pathogen Primers and Probes Design – mRNA In silico validation – mRNA Design and in silico validation-pathogen Multiplex PCR What can go wrong? Design principles and comparisons Reagents & Instrumentation MIQE Web-based resources
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