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Quantitative PCR Protocols

Handbook / ManualOut of Print

Series: Methods in Molecular Biology

Edited By: Bernd Kochanowski and Udo Reischl

304 pages, Figs, tabs

Humana Press

Hardback | Dec 1999 | #86821 | ISBN: 0896035182
Out of Print Details

About this book

This is 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. With its step-by-step instructions, Quantitative PCR Protocols allows researchers to address biological and diagnostic questions that are difficult or impossible to answer using any other experimental approach.

The book is very comprehensive and well written - The Biochemist


Contents

Part I. Reviews. Quantitative PCR: A Survey of the Present Technology, General Principles of Quantitative PCR, Effects of Collection, Processing, and Storage on RNA Detection and Quantification, Quantitative RT-PCR, Kinetic Quantitative PCR vs End-Point Quantitative PCR with Internal Standard, Comparison of Competitive PCR and Positive Control-Based PCR, Part II. Protocols. End-Point Titration-PCR for Quantitation of Cytomegalovirus DNA, Analysis of Amplified DNA Molecules by Capillary Electrophoresis and Laser Induced Fluorescence, Competitor Calibration and Analysis of Competitive Amplified PCR Products by High-Performance Liquid Chromatography (HPLC), Quantifying Amplicons with ELISA, Competitive PCR Quantitation Utilizing a Microtiter Plate Based Format for the Detection of PCR Products, Competitive and Differential RT-PCR (CD-RT-PCR) for Measurement of Normalized Gene Expression Using Antisense Competitors, Amplified Assay for Specific Dual-Labeled DNA Using the Coagulation Cascade (EDNA-ELCA), Quantitative PCR with Internal Standardization and OLA-ELISA Product Analysis for the p53 Tumor Suppressor Gene, Quantitative Analysis of Human DNA Sequences by PCR and Solid-Phase Minisequencing, High Resolution PCR Quantitation by AmpliSensor Assay, Construction of Polycompetitors for Competitive PCR, Tailed RT-PCR for the Quantitation of Chloramphenicol Acetyl Transferase (CAT)mRNA, A Stochastic PCR Approach for RNA Quantification in Multiple Samples, Quantitation of mRNA Species by RT-PCR on Total mRNA Population with Nonradioactive Probes, Index.

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