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.




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




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.




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




Plant Molecular Biology Manual


Book Description

Five years ago, the first edition of the Plant Molecular Biology Manual appeared. At that time, the editors felt that the field of plant molecular biology had matured to a point that the publication of a series of protocols in plant molecular biology was warranted. During the past five years, the field of plant molecular biology has expanded rapidly. This expansion is, among other things, reflected by the presence of several journals in the plant sciences, as well as by the increasing amount of plant sciences articles that are published in the more general journals. In 1991 approximately 3000 people attended the Third International Congress of Plant Molecular Biology in Tucson, Arizona, where more than 2000 posters were presented. It is also remarkable to see that nowadays botanical and physiological meetings pay a considerable amount of attention to plant molecular biology. Since the first edition of this manual appeared, we have published, yearly, a series of supplements to the original volume. These supplements covered new subjects and described new methods that had been developed. With time, however, the editors realized that the original manual plus supplements had become cumbersome to use, and we decided to publish a reorganized version of the manual.




Quantitation of mRNA by Polymerase Chain Reaction


Book Description

In this laboratory "cook-book", the authors provide a concise guide to PCR-based techniques to quantify nucleic acids in biological and clinical samples using exclusively nonradioactive detection methods, e.g. HPLC, biotin and digoxigenin based protocols. Each method presentation also includes sections on theory, reagents, standards, applicability, limitations, and trouble shooting. In addition to the protocols, the authors also provide the necessary information on: general aspects of nucleic acid quantitation; design of PCR standards; mRNA purification; cDNA synthesis; solution hybridization; DNA sequencing. This laboratory guide enables professionals as well as beginners to adopt easily quantitative PCR protocols into their own clinical or biomedical research.




Fluorescence Microscopy and Fluorescent Probes


Book Description

Fluorescence microscopy images can be easily integrated into current video and computer image processing systems. People like visual observation; they like to watch a television or computer screen, and fluorescence techniques are thus becoming more and more popular. Since true in vivo experiments are simple to perform, samples can be directly seen and there is always the possibility of manipulating the samples during the experiments; it is an ideal technique for biology and medicine. Images are obtained by a classical (now called wide-field) fluorescence microscope, a confocal scanning microscope, upright or inverted, with epifluorescence or transmission. Computerized image processing may improve definition, and remove glare and scattered light signal. It also makes it possible to compute ratio images (ratio imaging both in excitation and in emission) or lifetime imaging. Image analysis programs may supply a great deal of additional data of various types, starting with calculations of the number of fluorescent objects, their shapes, brightness, etc. Fluorescence microscopy data may be complemented by classical measurement in the cuvette yr by flow cytometry.




Basic Techniques in Molecular Biology


Book Description

This laboratory manual gives a thorough introduction to basic techniques. It is the result of practical experience, with each protocol having been used extensively in undergraduate courses or tested in the authors laboratory. In addition to detailed protocols and practical notes, each technique includes an overview of its general importance, the time and expense involved in its application and a description of the theoretical mechanisms of each step. This enables users to design their own modifications or to adapt the method to different systems. Surzycki has been holding undergraduate courses and workshops for many years, during which time he has extensively modified and refined the techniques described here.




Gene Transfer to Plants


Book Description




Drosophila melanogaster: Practical Uses in Cell and Molecular Biology


Book Description

Drosophila melanogaster: Practical Uses in Cell and Molecular Biology is a compendium of mostly short technical chapters designed to provide state-of-the art methods to the broad community of cell biologists, and to put molecular and cell biological studies of flies into perspective. The book makes the baroque aspects of genetic nomenclature and procedure accessible to cell biologists. It also contains a wealth of technical information for beginning or advanced Drosophila workers. Chapters, written within a year of publication, make this topical volume a valuable laboratory guide today and an excellent general reference for the future.Key Features* Collection of ready-to-use, state-of-the art methods for modern cell biological and related research using Drosophila melanogaster* Accessible to both experienced Drosophila researchers and to others who wish to join in at the cutting edge of this system * Drosophila offers an easily managed life cycle, inexpensive lifestyle, extraordinarily manipulable molecular and classical genetics, now combined with powerful new cell biology techniques * Introduction and overview sections orient the user to the Drosophila literature and lore * Six full-color plates and over 100 figures and tables enhance the understanding of these cell biology techniques