Quantitative PCR Protocols (Methods in Molecular Medicine) - Hardcover

 
9780896035188: Quantitative PCR Protocols (Methods in Molecular Medicine)

Synopsis

Since the polymerase chain reaction (PCR) was first developed in 1985, an enormous number of research reports have documented the versatility of this brilliant technique for in vitro amplification of nucleic acids. Although PCR has had a profound impact in many areas of research, contrary to exp- tation its routine application to the quantitation of nucleic acids has proven problematic in several aspects. The shortcomings are principally caused by the exponential nature of PCR, whereby small variations in amplification ef- ciency may dramatically affect the yield of amplification product. Even mi- mal temperature deviations that occur between adjacent wells of a thermocycler or day-to-day variations in the efficiency of nucleic acid preparation can lead to significant differences in the extent of amplification between otherwise id- tical samples. However, knowing more about the intrinsic limitations of PCR is the first step towards surmounting the shortcomings associated with this prom- ing methodology. With the introduction of appropriate standards of known amount, which are co-amplified with the sample using the same primers, it is increasingly feasible to address biological or diagnostic questions that are d- ficult or impossible to answer using any other experimental approach.

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From the Back Cover

In Quantitative PCR Protocols, Bernd Kochanowski and Udo Reischl assemble a significant collection of established and novel methods for the successful quantitation of nucleic acids. Each method has been refined and tested by its developer and proven to work in such problems as the analysis of eukaryotic gene expression, the quantitation of viral loads in clinical specimens, reporter gene expression, and quantitative oncogene analysis. Particular emphasis is placed on the underlying principles of the design of competitive or noncompetitive standards, as well as on the optimization of the amplification process. In important cases several methods are given for the same problem so that readers may set up test systems tailored to their specific practical needs.
Quantitative PCR Protocols provides a collection of detailed methods that offers comprehensive and up-to-date information on how to control PCR to obtain reliable quantitation. With its step-by-step instructions researchers can really master the complexities of PCR and address otherwise inaccessible biological and diagnostic questions.

Review

"...The book is very comprehensive and well written..."-The Biochemist

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