Molecular Biomethods Handbook

Recent advances in the biosciences have led to a range of powerful new technologies, particularly nucleic acid, protein and cell-based methodologies. The most recent insights have come to affect how scientists investigate and define cellular processes at the molecular level. This book expands upon the techniques included in the first edition, providing theory, outlines of practical procedures, and applications for a range of techniques. Written by a well-established panel of research scientists, the book provides an up-to-date collection of methods used regularly in the authors’ own research programs.

John Walker and Ralph Rapley have collected a wide-ranging group of molecular and biochemical techniques that are the most frequently used in medical and clinical research, especially diagnostics. The authors-well-established investigators who run their own research programs and use the methods on a regular basis-outline the practical procedures for using them and describe a variety of pertinent applications. Among the technologies presented are southern and western blotting, electrophoresis, PCR, cDNA and protein microarrays, liquid chromatography, in situ hybridization, karyotyping, flow cytometry, bioinformatics, genomics, and ribotyping. The applications include assays for mutation detection, mRNA analysis, chromosome translocations, inborn errors of metabolism, protein therapeutics, and gene therapy.

Recent advances in the biosciences have led to a range of powerful new technologies, particularly nucleic acid, protein and cell-based methodologies. The most recent insights have come to affect how scientists investigate and define cellular processes at the molecular level. This book expands upon the techniques included in the first edition, providing theory, outlines of practical procedures, and applications for a range of techniques. Written by a well-established panel of research scientists, the book provides an up-to-date collection of methods used regularly in the authors’ own research programs.

Since the publication of the best-selling Handbook of Molecular and Cellular Methods in Biology and Medicine, the field of biology has experienced several milestones. Genome sequencing of higher eukaryotes has progressed at an unprecedented speed. Starting with baker's yeast (Saccharomyces cerevisiae), organisms sequenced now include human (Homo sapiens), model crucifer (Arabidopsis thaliana), and rice (Oryza sativa). The invention of DNA microarray technology and advances in bioinformatics have generated vast amounts of genomic data. Reflecting these revolutionary advances Handbook of Molecular and Cellular Methods in Biology and Medicine, Second Edition documents conventional and modern approaches to tackle scientific research in the post-genomics era. Maintaining the step-by-step format that popularized the first edition, each chapter provides the principles behind the featured method, a detailed description of each protocol, applications of the protocol to different systems, and references for further study. Handbook of Molecular and Cellular Methods in Biology and Medicine, Second Edition now includes: New protocols in all chapters, including alternative protocols In vitro transcription methods Analysis of DNA sequences New bioseparation techniques New chapters covering: mRNA differential display Inhibition of gene expression In situ hybridization (Localization of gene expression) Combinatorial techniques Computational data mining methods applied to combinatorial chemistry libraries With this book at hand, researchers, teachers, and students can understand and utilize the major techniques and methods currently employed in cellular and molecular biology.

This laboratory guide comes at a time when several other method books have already been published in this field. Is this one different from the others? Yes and no. There was no attempt made to be comprehensive. Rather, data were brought to bear on areas where enough competence has been gathered in our laboratories and to complement recent method books (many of which cover extensively various aspects of molecular biology) in those matters which appeared to us somewhat neglected. There was a constant preoccupation and effort to provide miniaturized proce dures that are both simple and time-saving. Interest was devoted to standardized procedures and culture conditions, avoiding dogmas such as those giving excessive importance to sophisticated culture media with endless adjustments for local or personal considerations. The key to success is the quality of the plant material serving as a source of cells. Consequently, isolation. extraction or culture techniques can be simplified and standardized. This is symptomatic for our times as it marks the end of a period when methodological matters were frequently above the biological problems. The times of "methods above all" is basically over, despite the fact that many of us still believe that, say, tissue culture is a "science" per se. By presenting a few original techniques we believe that one seriously reduces the empiricism still prevailing in this area of research.

A comprehensive treasury of all the key molecular biology methods-ranging from DNA extraction to gene localization in situ-needed to function effectively in the modern laboratory. Each of the 120 highly successful techniques follows the format of the much acclaimed Methods in Molecular BiologyTM series, providing an introduction to the scientific basis of each technique, a complete listing of all the necessary materials and reagents, and clear step-by-step instruction to permit error-free execution. Included for each technique are notes about pitfalls to avoid, troubleshooting tips, alternate methods, and explanations of the reasons for certain steps-all key elements contributing significantly to success or failure in the lab. The Nucleic Acid Protocols Handbook constitutes today's most comprehensive collection of all the key classic and cutting-edge techniques for the successful isolation, analysis, and manipulation of nucleic acids by both experienced researchers and those new to the field.