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Since the publication of Nonistopic DNA Probe Techniques in 1992, the move away from radioactive materials for research and diagnostics has continued. This is due in part to public awareness of the hazards of radioactive waste and laws making radioactive disposal more difficult and costly and to improvement in both the sensitivity and convenience of nonisotopic techniques. Several new nonisotopic techniques have been developed and substantial improvements made to existing nonisotopic methods since 1992, and these are now included in Nonisotopic Probing, Blotting, and Sequencing. Nonisotopic Probing, Blotting, and Sequencing is an updated, expanded edition of the bestseller, Nonisotopic DNA Probe Techniques. It has been thoroughly revised to include the latest improvements in nonisotopic tagging techniques for macromolecules. Like its predecessor, it enables researchers to select the best nonisotopic method for their needs and maximize success by following its straightforward protocols. - Provides strategies and detailed procedures for labeling, blotting, and probing specific nucleic acid sequences and, with this edition, protein molecules - Gives protocols for nonisotopic DNA sequencing - new in this edition - Gives extensive, practical information - Presents background information for each method - Provides expert accounts from the inventor or developer of each method - Contains seven entirely new chapters - Covers all major types of nonisotopic procedures for labeling and detection
Biological Techniques is a series of volumes aimed at introducing to a wide audience the latest advances in methodology. The pitfalls and problems of new techniques are given due consideration, as are those small but vital details not always explicit in the methods sections of journal papers.In recent years, most biological laboratories have been invaded by computers and a wealth of new DNA technology and this will be reflected in many of the titles appearing in the series.The books will be of value to advanced researchers and graduate students seeking to learn and apply new techniques, and will be useful to teachers of advanced undergraduate courses involving practical or project work.Labelled biomolecules are an essential tool in life science research, and non-radioactive labels are becoming increasingly important due to their convenience of measurement, greater safety and lack of disposal problems compared to radioactive labels.This book provides practical information, background theory and protocols to allow a beginner to label many types of biomolecules, including proteins, peptides, nucleic acids and small molecules.This book is essential for biochemists, molecular biologists and cell biologists wanting to use non-radioactively labelled molecules.Aimed at researchers without specific expertise in chemistry, the book includes: - A review of the main signal systems and labels available, indicating their strengths and weaknesses - Discussion of the most useful strategies for labelling the various biomolecules - 32 protocols covering common labelling needs - Descriptions of the factors governing protocol design, enabling protocols to be modified for different applications - Sources of information including references, data and suppliers
New techniques and updated protocols for the detection and analysis of biomolecules - proteins, glycoproteins and nucleic acids. The second edition of this successful laboratory manual describes in detail the highly sensitive systems which are widely used in molecular biological and biomedical laboratories, such as colorimetric, luminescence, fluorescence measuring using antibody/antigen binding or hybridisation as well as PCR amplification. The clearly structured step-by-step protocols with practical hints and a troubleshooting guide are complemented by chapters on the theoretical background and the application of the techniques, enabling scientists to plan, design and conduct the appropriate procedures.
Molecular Probes—Advances in Research and Application: 2013 Edition is a ScholarlyBrief™ that delivers timely, authoritative, comprehensive, and specialized information about ZZZAdditional Research in a concise format. The editors have built Molecular Probes—Advances in Research and Application: 2013 Edition on the vast information databases of ScholarlyNews.™ You can expect the information about ZZZAdditional Research in this book to be deeper than what you can access anywhere else, as well as consistently reliable, authoritative, informed, and relevant. The content of Molecular Probes—Advances in Research and Application: 2013 Edition has been produced by the world’s leading scientists, engineers, analysts, research institutions, and companies. All of the content is from peer-reviewed sources, and all of it is written, assembled, and edited by the editors at ScholarlyEditions™ and available exclusively from us. You now have a source you can cite with authority, confidence, and credibility. More information is available at http://www.ScholarlyEditions.com/.
`In the second edition of Principles I have attempted to maintain the emphasis on basics, while updating the examples to include more recent results from the literature. There is a new chapter providing an overview of extrinisic fluorophores. The discussion of timeresolved measurements has been expanded to two chapters. Quenching has also been expanded in two chapters. Energy transfer and anisotropy have each been expanded to three chapters. There is also a new chapter on fluorescence sensing. To enhance the usefulness of this book as a textbook, most chapters are followed by a set of problems. Sections which describe advanced topics are indicated as such, to allow these sections to be skipped in an introduction course. Glossaries are provided for commonly used acronyms and mathematical symbols. For those wanting additional informtion, the final appendix contains a list of recommended books which expand on various specialized topics.' from the author's Preface
This volume details the theories, mechanisms, technologies and trends for solving qualitative and quantitative problems in diverse areas of analytical research - emphasizing physicochemical principles. It focuses on deriving simpler and more extensive chemiluminescence (CL) detectors reflecting miniaturization trends, including narrow-bone and capillary liquid chromatography versus high-performance liquid chromatography and miniaturized high-performance thin-layer chromatography. It also covers the sensitivity, selectivity, wide detection range and versatility of CL-based methodologies.
During the past 15 years, there has been remarkable progress in the analysis and manipulation of DNA and its use in nanotechnology. DNA analysis is ubiquitous in molecular biology, medical diagnostics, and forensics. Much of the readout technology is based on fluorescence detection. This volume contains contributions from many experts in the field who present an overview of many aspects of DNA technology. These chapters provide an understanding of the underlying principles and technology, rather than an exhaustive review of the literature. Written in a clear straightforward style, this book is an excellent introduction for any scientist to the use of fluorescence in DNA analysis. DNA Technology is an essential reading for all academics, bench scientists, and industry professionals wishing to take advantage of the latest and greatest in this continuously emerging field. Key Features: *Comprehensive overview of the complexities of DNA analysis, *Covers topics of universal interest to a broad field of scientists, *Accessible utility in presenting state-of-the-art DNA technology, *Chapters authored by key figures in the field.
The dangers and drawbacks inherent in radioactivity-based methods along with a demonstrated and dramatic increase in sensitivity have precipitated a major shift towards luminescence measurements and visualization techniques. Their use has now spread even to traditional clinical environments, and their applications have grown from clinical assays to
The new techniques of molecular cytogenetics, mainly fluorescence in situ hybridization (FISH) of DNA probes to metaphase chromosomes or interphase nuclei, have been developed in the past two decades. Many FISH techniques have been implemented for diagnostic services, whereas some others are mainly used for investigational purposes. Several hundreds of FISH probes and hybridization kits are now commercially available, and the list is growing rapidly. FISH has been widely used as a powerful diagnostic tool in many areas of medicine including pediatrics, medical genetics, maternal–fetal medicine, reproductive medicine, pathology, hematology, and oncology. Frequently, a physician may be puzzled by the variety of FISH techniques and wonder what test to order. It is not uncommon that a sample is referred to a laboratory for FISH without indicating a specific test. On the other hand, a cytogeneticist or a technologist in a laboratory needs, from case to case, to determine which procedure to perform and which probe to use for an informative result. To obtain the best results, one must use the right DNA probes and have reliable protocols and measures of quality assurance in place. Also, one must have sufficient knowledge in both traditional and molecular cytogenetics, as well as the particular areas of medicine for which the test is used in order to appropriately interpret the FISH results, and to correlate them with clinical diagnosis, treatment, and prognosis.
Until the mid 1980s, the detection and quantification of a specific mRNA was a difficult task, usually only undertaken by a skilled molecular biologist. With the advent of PCR, it became possible to amplify specific mRNA, after first converting the mRNA to cDNA via reverse transcriptase. The arrival of this technique—termed reverse transcription-PCR (RT-PCR)—meant that mRNA suddenly became amenable to rapid and sensitive analysis, without the need for advanced training in molecular biology. This new accessibility of mRNA, which has been facilitated by the rapid accumulation of sequence data for human mRNAs, means that every biomedical researcher can now include measurement of specific mRNA expression as a routine component of his/her research plans. In view of the ubiquity of the use of standard RT-PCR, the main objective of RT-PCR Protocols is essentially to provide novel, useful applications of RT-PCR. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the basic RT-PCR protocol. For example, a variety of different adaptations are described that have been employed to obtain quantitative data from RT-PCR. Quantitative RT-PCR provides the ability to accurately measure changes/imb- ances in specific mRNA expression between normal and diseased tissues.