Nonisotopic Probing, Blotting, and Sequencing


Book Description

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




Protocols for Nucleic Acid Analysis by Nonradioactive Probes


Book Description

Protocols for Nucleic Acid Analysis by Non-radioactive Probes, Second Edition provides a firm background on the basic preparative protocols required for the analysis of nucleic acids by nonradioactive methods. Presenting the methodologies using amazing new applications, this volume offers guide chapters on nucleic acid extractions, preparation of nucleic acid blots, and labeling of nucleic acids with nonradioactive haptens. New fluorescent techniques such as Real Time PCR and microarrays are also included, allowing users to get a nonradioactive protocol implemented in the laboratory with minimum adaptation required and fastest time to results. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.




Nonradioactive Labeling and Detection of Biomolecules


Book Description

All the bioanalytical labeling and detection techniques in one source! This book gathers together all the important nonradioactive labeling techniques for nucleic acids, proteins, glycoproteins and glycolipids like Digoxigenin:Anti-Dioxigenin (DIG), Biotin, 5-Bromodeoxyuridine (BrdU), Sulfone, Immunogold, Silver Enhancement, and Synthetic Nucleic Acid Probe (SNAP) as well as the standard procedures for optical, chemical, biological, electrochemiluminescent and fluorescent detection. Additionally, applications for the use of non-isotopically labeled biomolecules are described. Specific protocols are given for hybridization analysis such as blot, colony/plaque and in-situ hybridization formats, quantitative formats, and also nonradioactivetechniques for nucleic acid sequencing and amplification. Each chapter contains a short introduction, a detailed description of the method with labprotocols, troubleshooting tips and references.




Nonradioactive Analysis of Biomolecules


Book Description

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.




Transgenic Plants


Book Description

The aim of Transgenic Plants: Methods and Protocols is to provide a source of information to guide the reader through a wide range of frequently used, broadly applicable, and easily reproducible techniques involved in the gene- tion of transgenic plants. Its step-by-step approach covers a series of methods for genetically transforming plant cells and tissues, and for recovering whole transgenic plants from them. The volume then moves on to the use of sele- able and reporter markers, positive selection, marker elimination after rec- ery of transgenic plants, and the analysis of transgene integration, expression, and localization in the plant genome. Although contributors usually refer to model plants in most chapters, the protocols described herein should be widely applicable to many plant species. The last two sections are devoted to me- ods of risk assessment and to exploring the current and future applications of transgenic technology in agriculture and its social implications in a case study. Transgenic Plants: Methods and Protocols is divided into six major s- tions plus an introduction, comprising 27 chapters. Part I, the Introduction, is a review of the past, present, and perspectives of the transgenic plants, from the discovery of Agrobacterium tumefaciens as a feasible transformation vector, to its use as a tool to study gene expression and function, and the current and possible future applications of this technology in agriculture, industry, and medicine.




Non-Radioactive Labelling


Book Description

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




Rapid Methods and Automation in Microbiology and Immunology


Book Description

The current diagnostic methods for the great variety of microbial agents affecting health are clearly unsatisfactory. New important pathogens have emerged including the agent responsible for bovine spongiform encephalopathy. Moreover, there is an increasing need for more accurate microbial control of our environment, and of the food and water we consume. What is needed are rapid, sensitive and reliable procedures which, on the one hand, should be suitable for automation and, onthe other hand, presented in a cost-effective version suitable for field use. Including new biochemical approaches, such as polymerase chain reaction, recombinant gene products and synthetic peptides, these needs are discussed in these protocols of the RAMI-90 congress.







Nonisotopic DNA Probe Techniques


Book Description

Recently many nonisotopic methods of probing specific DNA sequences have been developed as replacements for radioactive labels, such as 32phosphorous and 125iodine. This book brings all of these new methods together in one convenient, easily accessible source. It enables researchers to select the nonisotopic method best suited to their application and to use it to maximum advantage by following the straightforward instructions provided.This book contains chapters on colorimetric, bioluminescent, chemiluminescent, fluorescent, and time-resolved fluorescent detection methods. Each chapter has been written by the inventor or developer of a particular nonisotopic method and thus provides an expert account of the method. Each chapterpresents useful background information and detailed, step-by-step, easy-to-follow, experimental procedures for labeling and detection.Gives extensive practical informationCovers major types of nonisotopic labels and proceduresPresents background information for each methodProvides strategies and detailed experimental procedures for labeling and detecting DNA sequences byFluorescenceChemiluminescenceBioluminescenceColorimetry