Search results for: cdna-library-protocols

Cdna Library Protocols

Author : Ian G. Cowell
File Size : 66.16 MB
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The first libraries of complementary DNA (cDNA) clones were con structed in the mid-to-late 1970s using RNA-dependent DNA polymerase (reverse transcriptase) to convert poly A* mRNA into double-stranded cDNA suitable for insertion into prokaryotic vectors. Since then cDNA technology has become a fundamental tool for the molecular biologist and at the same time some very significant advances have occurred in the methods for con structing and screening cDNA libraries. It is not the aim of cDNA Library Protocols to give a comprehensive review of all cDNA library-based methodologies; instead we present a series of up-to-date protocols that together should give a good grounding of proce dures associated with the construction and use of cDNA libraries. In deciding what to include, we endeavored to combine up-to-date versions of some of the most widely used protocols with some very usefiil newer techniques. cDNA Library Protocols should therefore be especially useful to the investigator who is new to the use of cDNA libraries, but should also be of value to the more experienced worker. Chapters 1--5 concentrate on cDNA library construction and manipula tion, Chapters 6 and 7 describe means of cloning difficult-to-obtain ends of cDNAs, Chapters 8-18 give various approaches to the screening of cDNA libraries, and the remaining chapters present methods of analysis of cDNA clones including details of how to analyze cDNA sequence data and how to make use of the wealth of cDNA data emerging from the human genome project.

CDNA Library Protocols Methods in Molecular Biology

Author : Ian G. Cowell
File Size : 80.22 MB
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This comprehensive collection of detailed protocols covers all areas of cDNA work, from library construction and manipulation to screening and analysis of resulting clones. Great care has been taken to combine up-to-date versions of some of the most widely used protocols with some very useful newer techniques. The protocols describe methods for cloning difficult-to-obtain ends of cDNAs, methods for analyzing cDNA sequence data, and methods for using the wealth of cDNA data emerging from the human genome project. Bearing in mind the importance of the library screening method to the determination of cloning strategy, the book offers a wide range of approaches to screening cDNA libraries.

Generation of CDNA Libraries

Author : Shao-Yao Ying
File Size : 24.42 MB
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Expert researchers and inventors in the field describe their own proven techniques for generating cDNA/mRNA libraries to identify the functions of specific decoded gene sequences. A wide variety of techniques is presented for enhancing the generation of complete and full-length libraries, and for confirming the quality of the cDNAs generated. Among the applications detailed are electrophoresis, Northern blotting, single cell microarray analysis, subtractive hybridization, subtractive cloning, gene cloning, and peptide library generation.

Some Protocols for CDNA Library Construction with Conifer Tissues

Author : J. A. Pitel
File Size : 46.30 MB
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Yeast Protocols

Author : Wei Xiao
File Size : 61.24 MB
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Since the publication of Yeast Protocols in 1996, many new techniques have been invented and original protocols improved and refined. This throughly updated second edition will serve as a stand-alone protocols handbook of these new and refined techniques suitable for daily use in all research laboratories. It includes all of the recent advanced protocols as well as the major basic techniques and hence, will be essential for both yeast research laboratories and those researchers who wish to use yeast as a host to study their favorite genes from other organisms.

Malaria Methods and Protocols

Author : Denise L. Doolan
File Size : 83.94 MB
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Despite considerable scientific and medical effort over the past decades, malaria remains the most important human parasitic disease. It is responsible for up to 3 million deaths and another 300-500 million new cases each year, and is becoming resistant to the current chemoprophylactic and chemotherapeutic agents. In Malaria Methods and Protocols, internationally respected scientists and clinicians describe in step-by-step detail their most useful conventional and cutting-edge techniques for the study of malaria. Areas covered include clinical and laboratory diagnosis and typing, animal models, molecular biology, immunology, cell biology, vaccinology, laboratory models, and field applications. Each readily reproducible protocol has been tested, standardized, and optimized for experimental success, and includes many laboratory notes on troubleshooting, avoiding pitfalls, and interpreting results. Several of the most widely used methods are either described here in detail for the first time or have been thoroughly updated since their original publication (e.g., in vitro culture of Plasmodium parasites and in vitro growth inhibition assay). State-of-the-art and highly practical, Malaria Methods and Protocols makes available to basic and applied researchers today's only comprehensive collection of essential laboratory methods for diagnosing malaria, characterizing the parasite, understanding the interaction between the human host and Plasmodium parasite, and developing effective preventive measures.

Reviews and Protocols in DT40 Research

Author : Jean-Marie Buerstedde
File Size : 20.45 MB
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The vertebrate genome DT40 has proven to be a reliable and robust research subject, with fast doubling time, easy clonability and a relatively stable karyotype. This book provides an up to date overview of the different facets of research, and also intends to help newcomers get started and avoid looming pitfalls. The collection of protocols which have been provided by a number of laboratories will be particularly useful in this regard.

Surgical Research

Author : Wiley W. Souba
File Size : 85.81 MB
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Contributors. -- Foreword. -- Preface. -- Getting Started. -- Assessing Available Information. -- Organizing and Preliminary Planning for Surgical Research -- Writing a Protocol: Animals, Humans, and Use of Biologic, Chemical, and Radiologic Agents. -- Grantsmanship. -- Informed Consent and the Protection of Human Research Subjects: Historical Perspectives and Guide to Current United States Regulations. -- Animal Care and Maintenance. -- Funding Strategies and Agencies: Academic-Industrial Relationships; Intellectual Property. -- Statistical Considerations. -- Use of Nonexperimental Studies to Evaluate Surgical Procedures and Other Interventions: The Challenge of Risk Adjustment. -- Measuring Surgical Outcomes. -- Design of Clinical Trials. -- Using Administrative Data for Clinical Research. -- Research in the Intensive Care Unit: Ethical and Methodological Issues. -- Research in the Operating Room. -- Effects of Age and Gender. -- Strategies, Principles, and Techniques Using Transgeni ...

The Nucleic Acid Protocols Handbook

Author : Ralph Rapley
File Size : 77.89 MB
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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 BiologyOao 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."

Natural Killer Cell Protocols

Author : Kerry S. Campbell
File Size : 49.20 MB
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In Natural Killer Cell Protocols: Cellular and Molecular Methods, Kerry S. Campbell and Marco Colonna have assembled a comprehensive collection of readily reproducible methods designed to study natural killer (NK) cells from the broadest variety of viewpoints. These include not only classic techniques, but also new approaches to standard methods, newly evolved techniques that have become valuable for specific applications, and unique models for manipulating and studying NK cells. Among the advanced methods covered are those for in vitro transendothelial migration, in vivo detection of cells migrating into tumors, immunofluorescence staining of intracellular cytokines, and in vitro NK cell development. Valuable techniques for specific applications include vaccinia virus protein expression, soluble KIR-Fc fusions for HLA class I binding assays, calcium mobilization in cell conjugates, and identification of heterodimeric receptor complexes using cDNA library expression cloning. No less important are accounts of such classic methods as hybrid resistance, ADCC, viral defense, target cell cytotoxicity assays, cloning and culturing, tumor immunotherapy, and generation of HLA class I transfected target cells. Natural Killer Cell Protocols: Cellular and Molecular Methods offers immunologists, cancer researchers, virologists, and cell biologists today's most comprehensive collection of both established and cutting-edge techniques, methods that will contribute significantly to advancing our understanding of this fascinating and critically important class of cells.