Protocols For Nucleic Acid Analysis By Nonradioactive Probes

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.

With each chapter written by a distinguished expert in the biochemistry field, this comprehensive volume describes the preparation and use of a variety of radioactive and non-radioactive probes in situations ranging from research laboratories to routine diagnosis laboratories. The enzymatic and chemical techniques for labeling nucleic acid probes with radioisotopes and with non-radioactive ligands and haptens are discussed. Additionally, the associated methods for their detection, the use of these probes in the diagnosis of human and microbial pathogens, of plant viruses and viroids, and of human genetic disorders, as well as in the detection of nucleic acids in tissues and cells by in situ hybridization are presented.

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.

This edition is an updated version of the first edition, plus many new sections on specific applications of nucleic acid probes. Due to the explosion of such applications, experts in each major area have described their specialty. As in the first edition, the emphasis is on the commercial uses of DNA probes, as in diagnostic applications, and it incorporates background material, advice and specific protocols.

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 information Covers major types of nonisotopic labels and procedures Presents background information for each method Provides strategies and detailed experimental procedures for labeling and detecting DNA sequences by Fluorescence Chemiluminescence Bioluminescence Colorimetry

An authoritative team of investigators illuminate the core bioanalytical techniques used every day in their own laboratories, and laboratories throughout the world. These highly experienced scientists fully explain both the theory behind, and the application of, these key techniques, and include extensive references for those seeking detailed laboratory protocols. The techniques covered range from the extraction, separation, detection, and characterization of nucleic acids to gene cloning and library production, mapping, expression, transgenesis, differential display, and DNA profiling, to name a few. Numerous key protein methods, as well as support and related techniques, are also included. The goal is to provide established scientists and novices who are new to these techniques with a deeper understanding of the widest variety of biotechniques and their applications.

In Nucleic Acid Chemistry: Methods and Protocols, expert researches in the field detail techniques and approaches for the detection of DNA and RNA. These techniques include the recovery of trace amounts of DNA for amplification and analysis, new qPCR chemistries, new application of isothermal amplification techniques, assays with visual or electric signals for point-of-care diagnostics, improvement of fluorescent in situ hybridization, and new signal amplification techniques. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Nucleic Acid Chemistry: Methods and Protocols seeks to aid scientists in the further study of detection for DNA and RNA.

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