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Basic Methods in Molecular Biology discusses the heart of the most recent revolution in biology—the development of the technology of genetics. The achievements in this field have simply changed what biologists do and, perhaps even more important, the way they think. Moreover, never before have scientists from such a broad range of disciplines rushed into such a small and slightly arcane field to learn and carry off a bit of the technology. This book comprises 21 chapters, opening with three introductory ones that discuss the basics of molecular biology; the tools of the molecular biologist; and general preparations, procedures, and considerations for use of the book. The following chapters then discuss cloning vectors and bacterial cells; preparation of DNA from eukaryotic cells; probing nucleic acids; plasmid DNA preparation; DNA restriction fragment preparation; purification of DNA; and preparation and analysis of RNA from eukaryotic cells. Other chapters cover preparation of DNA from bacteriophage clones; cloning DNA from the eukaryotic genome; subcloning into plasmids; M13 cloning and sequencing; further characterization of cloned DNA; transfection of mammalian cells in culture; protein methods; general methods; and specialized methods. This book will be of interest to practitioners in the fields of biology and molecular genetics.
This manual not only provides reliable, up-to-date protocols for lab use but also the theoretical background of molecular biology, allowing users to better understand the principles underlying these techniques. It covers a wide range of methods, including the purification of nucleic acids, enzymatic modification of DNA, isolation of specific DNA fragments, PCR, cloning techniques, and gene expression. A Springer Lab Manual
This volume has evolved from a laboratory methods book that one of us first compiled nearly fifteen years ago. Since that time the book has undergone many minor revisions in order to include new methods and updated versions of older methods. The result has been an increasingly useful and more widely circulated book. However, the recent series of technological explosions generally lumped together under the name of "recombinant DNA technology" has been a turning point in the evolution of this previously underground publication. Minor revisions will no longer do. To keep the book useful we have had to make major revi sions and additions. The result is a dramatically expanded book that should be more useful to more people. The larger size and wider usefulness of the book have made this more formal publication seem a reasonable step to take. One of the reasons that this volume should be useful to many people is that it includes only procedures that have been used repeatedly by us and that have proven highly reliable both to ourselves and to others in our laboratories.
Current techniques for studying biological macromolecules and their interactions are based on the application of physical methods, ranging from classical thermodynamics to more recently developed techniques for the detection and manipulation of single molecules. Reflecting the advances made in biophysics research over the past decade, and now including a new section on medical imaging, this new edition describes the physical methods used in modern biology. All key techniques are covered, including mass spectrometry, hydrodynamics, microscopy and imaging, diffraction and spectroscopy, electron microscopy, molecular dynamics simulations and nuclear magnetic resonance. Each method is explained in detail using examples of real-world applications. Short asides are provided throughout to ensure that explanations are accessible to life scientists, physicists and those with medical backgrounds. The book remains an unparalleled and comprehensive resource for graduate students of biophysics and medical physics in science and medical schools, as well as for research scientists looking for an introduction to techniques from across this interdisciplinary field.
Developmental biology is one of the most exciting and fast-growing fields today. In part, this is so because the subject matter deals with the innately fascinating biological events—changes in form, structure, and function of the org- ism. The other reason for much of the excitement in developmental biology is that the field has truly become the unifying melting pot of biology, and provides a framework that integrates anatomy, physiology, genetics, biochemistry, and cellular and mole- lar biology, as well as evolutionary biology. No longer is the study of embryonic development merely “embryology.” In fact, development biology has produced - portant paradigms for both basic and clinical biomedical sciences alike. Although modern developmental biology has its roots in “experimental emb- ology” and the even more classical “chemical embryology,” the recent explosive and remarkable advances in developmental biology are critically linked to the advent of the “cellular and molecular biology revolution.” The impressive arsenal of expe- mental and analytical tools derived from cell and molecular biology, which promise to continue to expand, together with the exponentially developing sophistication in fu- tional imaging and information technologies, guarantee that the study of the devel- ing embryo will contribute one of the most captivating areas of biological research in the next millennium.
The last few years have seen the rapid development of new methodology in the field of molecular biology. New techniques have been regularly intro duced and the sensitivity of older techniques greatly improved upon. Developments in the field of genetic engineering in particular have con tributed a wide range of new techniques. In Volume 1, published in 1983, we introduced the reader to a selection of the more advanced analytical and preparative techniques which we considered to be frequently used by research workers in the field of molecular biology. In choosing techniques for Volume 1 we obviously had to be selective and were unable to cover as broad a spectrum of techniques as we would have liked. However, the pro duction of Volume 2 has allowed us to develop the theme initiated in Volume 1 and also expand to include a wider range of subject areas. As with Volume 1, the majority of chapters relate to nucleic acid method ology, but we have also covered immunological methodology and protein 1. Obviously, we purification techniques that were not included in Volume see Volume 2 as simply a continuation of Volume 1. As with Volume 1, a knowledge of certain basic biochemical techniques and terminology has been assumed. However, since many areas of molecular biology are developing at a formidable rate and constantly generating new termin ology, a glossary of terms has been included.
B-lymphocyte development and function remains an exciting area of research for those interested in the physiology and pathology of the immune system in higher animals. While recent advances in genetics and cellular and molecular biology have provided a large spectrum of powerful new experimental tools in this field, it is both time consuming and often very difficult for a student or just any bench-side worker to identify a reliable experimental protocol in the ocean of the literature. The aim of B Cell Protocols is to provide a collection of diverse protocols ranging from the latest inventions and applications to some classic, but still frequently used methods in B-cell biology. The authors of the various chapters are all highly qualified scientists who are either the inventors or expert users of these methods. Their extensive experience in mastering a particular method provides not only the step-by-step details of a reproducible protocol, but also useful troubleshooting tips that readers will appreciate in their daily work. We hope that this book will be helpful for both beginning and experienced researchers in the field in designing or modifying an experimental approach, and exploring a biological question from multiple angles.