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With a variety of detection chemistries, an increasing number of platforms, multiple choices for analytical methods and the jargon emerging along with these developments, real-time PCR is facing the risk of becoming an intimidating method, especially for beginners. Real-time PCR provides the basics, explains how they are exploited to run a real-time PCR assay, how the assays are run and where these assays are informative in real life. It addresses the most practical aspects of the techniques with the emphasis on 'how to do it in the laboratory'. Keeping with the spirit of the Advanced Methods Series, most chapters provide an experimental protocol as an example of a specific assay.
Geneticists and molecular biologists have been interested in quantifying genes and their products for many years and for various reasons (Bishop, 1974). Early molecular methods were based on molecular hybridization, and were devised shortly after Marmur and Doty (1961) first showed that denaturation of the double helix could be reversed - that the process of molecular reassociation was exquisitely sequence dependent. Gillespie and Spiegelman (1965) developed a way of using the method to titrate the number of copies of a probe within a target sequence in which the target sequence was fixed to a membrane support prior to hybridization with the probe - typically a RNA. Thus, this was a precurso...
Examines the latest innovations and the overall impact of PCR on areas of molecular research.
Quantitative Real-Time PCR: Methods and Protocols focuses on different applications of qPCR ranging from microbiological detections (both viral and bacterial) to pathological applications. Several chapters deal with quality issues which regard the quality of starting material, the knowledge of the minimal information required to both perform an assay and to set the experimental plan, while the others focus on translational medicine applications that are ordered following an approximate logical order of their medical application. The last part of the book gives you an idea of an emerging digital PCR technique that is a unique qPCR approach for measuring nucleic acid, particularly suited for l...
Rapid-Cycle Real-Time PCR is a powerful technique for nucleic acid amplification and analysis that often requires less than half an hour to perform. Samples are amplified by rapid-cycle PCR followed by immediate melting curve analysis in the same instrument. Melting curve analysis of PCR products with SYBR Green I often allows product identification without gel electrophoresis. Furthermore, in the presence of fluorescent hybridization probes, melting curves provide "dynamic dot blots" for fine sequence analysis, including single nucleotide polymorphisms (SNPs). The method is often cited as the most versatile, efficient method for nucleic acid analysis in research and diagnostics in the fields of genetics and oncology. Molecular diagnostics has never been easier!
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The 1st MOVE symposium in Málaga provided a highly prosperous event for interactions among international and young EV scientists from 16 European EV societies. Topics covered almost all fields of the current and potential future EV research areas, from methodological improvements, over fundamental biological EV topics, up to general physiological, disease, or cancer-related aspects.
A decade ago Leaf, a cancer survivor himself, began to investigate why we had made such limited progress fighting this terrifying disease. The result is a gripping narrative that reveals why the public's immense investment in research has been badly misspent, why scientists seldom collaborate and share their data, why new drugs are so expensive yet routinely fail, and why our best hope for progress-- brilliant young scientists-- are now abandoning the search for a cure.
Flavor science is continually evolving. Remaining current with the latest research and establishing a broad and sound understanding of the major developments and breakthroughs can be a challenge. The Weurman Flavour Research Symposium has long been regarded as the premier professional meeting focused on the science of flavor. Flavour Science, an extensive review of the most recent symposium, presents the latest in flavor research, enriching the chemistry-based vision of most flavorists and flavor chemists with understanding from a broad range of fields, including human physiology, ethology, psychophysics, genetics, bioinformatics or metabolomics. This book is ideal for all flavor scientists, food chemists and sensory scientists and has a strong audience among enologists as well. - Focuses on the rapidly changing field of flavor science - Includes the latest information on the physiology, chemistry and measurement of flavor - Presents practical information on the flavor industry and emerging trends
PCR’s simplicity as a molecular technique is, in some ways, responsible for the huge amount of innovation that surrounds it, as researchers continually think of new ways to tweak, adapt, and re-formulate concepts and applications. PCR Technology: Current Innovations, Third Edition is a collection of novel methods, insights, and points of view that provides a critical and timely reference point for anyone wishing to use this technology. Topics in this forward-thinking volume include: The purification and handling of PCR templates The effect of the manufacture and purification of the oligonucleotide on PCR behavior Optimum buffer composition Probe options The design and optimization of qPCR ...