BeschreibungThe Nucleic Acid Protocols Handbook is today's most comprehensive up-to-date 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 Biology(TM) series, providing an introduction to the scientific basis of each technique, a complete listing of all the materials and reagents needed, and clear step-by-step instructions 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 in the lab. The authord are well-experienced masters of the techniques described, often having perfected and optimized them in their own laboratories. In The Nucleic Acid Protocols Handbook, Ralph Rapley has brought together the widest-ranging, one-volume collection of classic and cutting-edge techniques for the successful isolation, analysis, and manipulation of nucleic acids. Eminently practical and reproducible, these proven methods will help both experienced researchers and those entering the field attain the laboratory mastery needed-not only to deepen understanding of biological processes at the molecular level, but also to begin realizing the biotechnological bounty so clearly in prospect.
InhaltsverzeichnisPart I. Nucleic Acid Extraction. Isolation of High-Molecular-Weight DNA from Animal Cells, Ian Garner. Isolation of mRNA by Affinity Chromatography, Sian Bryant and David L. Manning. Isolation and Purification of DNA from Plants, Justin Stacey and Peter G. Isaac. Purification of Uncontaminated, Intact Plant RNA, Shu-Hua Cheng, Brandon D. Moore, and Jeffrey R. Seemann. An Improved Method to Isolate Mitochondrial RNA from Green Plant Tissue, Fei Ye and Ralf Reski. Isolating Chromosomal DNA from Bacteria, Elisabeth Chachaty and Patrick Saulnier. Bacterial DNA Extraction for Polymerase Chain Reaction and Pulsed-Field Gel Electrophoresis, Elisabeth Chachaty and Patrick Saulnier. Isolation of Fungal Nucleic Acids, Surapareddy Sreenivasaprasad. Total RNA Isolation from Bacteria, John Heptinstall. Simultaneous RNA and DNA Extraction from Biopsy Material, Culture Cells, Plants, and Bacteria, Udo Döbbeling. Spectrophotometric Analysis of Nucleic Acids, John Heptinstall and Ralph Rapley. Part II. Basic Separation and Analysis of DNA. Restriction Endonuclease Digestion of DNA, Duncan R. Smith. Agarose Gel Electrophoresis of Nucleic Acids, D. Ross Williams and Ralph Rapley. Preparation of RNA Dot Blots, Rachel Hodge. Native Polyacrylamide Gel Electrophoresis, Adrian J. Harwood. Southern Blotting of Agarose Gels by Capillary Transfer, Ralph Rapley and Jane Davenport -Jones. Pulsed-Field Gel Electrophoresis, John Maule. HPLC of DNA and PCR Products, Elena D. Katz. Part III. Probe Design, Synthesis, and Labeling. End-Labeling of DNA Fragments, Adrian J. Harwood. Nick Translation and Random Hexamer Labeling of DNA, Jane Davenport-Jones. Generation of Labeled Probes by Polymerase Chain Reaction, Y. M. Dennis Lo and Shu F. An. Nonradioactive Oligonucleotide Probe Labeling, Sue Fowler and Ian Durrant. Preparation of Direct, Enzyme-Labeled DNA Probes, Ian Durrant and Timothy Stone. Random Prime Labeling of DNA Probes with Fluorescein-Tagged Nucleotides, Bronwen M. Harvey, Claire B.Wheeler, and Martin W. Cunningham. Hybridization and Detection of Fluorescein-Labeled DNA Probes Using Chemiluminescence, Claire B. Wheeler, Bronwen M. Harvey, and Martin W. Cunningham. Hybridization of Enzyme-Labeled Probes and Detection by Chemiluminescence, Timothy Stone and Ian Durrant. Hybridization and Competition Hybridization of Southern Blots, Rosemary Kelsell. Autoradiography and Fluorography, Eric Quéméneur. Part IV. RNA Analysis Techniques. Formaldehyde Gel Electrophoresis of Total RNA, Sian Bryant and David L. Manning. RNA Probes for the Analysis of Gene Expression, Dominique Belin. Primer Extension Analysis of mRNA, Maggie Walmsley, Mark Leonard, and Roger Patient. S1 Mapping Using Single-Stranded DNA Probes, Stéphane Viville and Roberto Mantovani. Measurements of Rate of Transcription in Isolated Nuclei by Nuclear 'Run-Off' Assay, Rai Ajit K. Srivastava and Gustav Schonfeld. One-Tube RT-PCR with Sequence-Specific RT Primers, Ulrich Pfeffer and Paola Ferro. Characterization of RNA Using Continuous RT-PCR Coupled with ELOSA, François Mallet. Quantitative Analysis of RNA Species by Polymerase Chain Reaction and Solid-Phase Minisequencing, Anu Suomalainen and Ann-Christine Syvänen. Nonradioactive Northern Blotting of RNA, Rainer Löw. Analysis of RNA by Northern Blotting Using Riboprobes, Rai Ajit K. Srivastava. Part V. Gene Library Construction and Screening. Production of Double-Stranded cDNA for Gene Library Synthesis, Jane Kirk and Steve Mayall. Using Rapid Amplification of cDNA Ends (RACE) to Obtain Full-Length cDNAs, Yue Zhang and Michael A. Frohman. cDNA Library Construction Using Streptavidin-Paramagnetic Beads and PCR, Kris N. Lambert and Valerie M. Williamson. Rapid (Ligase-Free) Subcloning of Polymerase Chain Reaction Products, Alan R. Shuldiner and Keith Tanner. Subtraction Hybridization cDNA Libraries, Clifford W. Schweinfest, Peter S. Nelson, Michael W. Graber, Rita I. Demopoulos, and Takis S. Papas. Cloning Polymerase Chain Reaction
Pressestimmen"... the techniques are presented in minute details and in a "cookbook-standardized manner"... The personal notes inserted by each contributor provide assurance that the techniques would work in any laboratory... Having all of those protocols updated and tested in practice and bound in one volume will be attractive to many scientists who deal with molecular biology techniques."- Modern Pathology
"There were over 190 authors involved in writing the protocols, ...each protocol is presented in outline format an follows a common numbering scheme, making it easy to find relevant sections....Many of the protocols include tables of data or figures, including drawings or photographs of gels...One of the most useful sections in each protocol lists notes, which elucidate possible pitfalls, provide troubleshooting tips, or give alternate methods or explanations for steps. This very practical information makes successful duplication of the procedure much more likely than when attempting to follow the often less detailed or less flexible descriptions given in journal articles....this handbook would be extremely useful as a reference in most university libraries."-E-Streams
"...it covers practically all of the very important and large field of nucleic acid research. The volume summarizes, in 10 parts, recent advances in the isolation , analysis and manipulation of nucleic acids. ...The chapters were contributed by authors from leading laboratories in the field all over the world....The book provides the reader with a large set of ready-to use protocols needed to undertake complex molecular biology experimental project on a large spectrum of biological models involving isolation and analysis of nucleic acids. It is thus an almost indispensable handbook for anybody working in the field of molecular biology and genetics."-Folia Microbiologica
"...extremely useful as a reference in most university libraries." - E-Streams
"...comprehensive and detailed...this is an set of recipes for the genetic kitchen." -The Biologist
Untertitel: 2000. Auflage. Sprache: Englisch.
Verlag: SPRINGER VERLAG GMBH
Erscheinungsdatum: März 2000
Seitenanzahl: 478 Seiten