Search results for: cdna-library-protocols

cDNA Library Protocols

Author : Ian G. Cowell
File Size : 83.52 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 : 36.9 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 : 77.93 MB
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Since its invention and subsequent development nearly 20 years ago, po- merase chain reaction (PCR) has been extensively utilized to identify numerous gene probes in vitro and in vivo. However, attempts to generate complete and full-length complementary cDNA libraries were, for the most part, fruitless and remained elusive until the last decade, when simple and rapid methods were developed. With current decoding and potential application of human genome information to genechips, there are urgent needs for identification of functional significance of these decoded gene sequences. Inherent in bringing these app- cations to fruition is the need to generate a complete and full-length cDNA library for potential functional assays of specific gene sequences. Generation of cDNA Libraries: Methods and Protocols serves as a laboratory manual on the evolution of generation of cDNA libraries, covering both ba- ground information and step-by-step practical laboratory recipes for which p- tocols, reagents, operational tips, instrumentation, and other requirements are detailed. The first chapter of the book is an overview of the basics of generating cDNA libraries, which include the following: (a) the definition of a cDNA library, (b) different kinds of cDNA libraries, (c) differences between methods for cDNA library generation using conventional approaches and novel stra- gies, including reverse generation of RNA repertoires from cDNA libraries, and (d) the quality of cDNA libraries.

Methods in Molecular Biology cDNA library protocols

Author : John M. Walker
File Size : 87.66 MB
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Yeast Protocols

Author : Wei Xiao
File Size : 86.79 MB
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In this second edition of a widely used classic laboratory manual, leading experts utilize the tremendous progress and technological advances that have occurred to create a completely new collection of not only the major basic techniques, but also advanced protocols for yeast research and for using yeast as a host to study genes from other organisms. The authors provide detailed methods for the isolation of subcellular components-including organelles and macromolecules, for the basic cellular and molecular analysis specific for yeast cells, and for the creation of conditional mutant phenotypes that lend themselves to powerful genome manipulation. Additional protocols offer advanced approaches to study genetic interactions, DNA and chromatin metabolism, gene expression, as well as the foreign genes and gene products in yeast cells.

PCR Cloning Protocols

Author : Bing-Yuan Chen
File Size : 87.10 MB
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PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.

Malaria Methods and Protocols

Author : Denise L. Doolan
File Size : 45.66 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.

RT PCR Protocols

Author : Joe O'Connell
File Size : 39.75 MB
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A panel of highly regarded molecular biologists and clinical researchers describe in detail their most novel, useful, and interesting RT-PCR applications. Here the newcomer will find readily reproducible protocols for highly sensitive detection and quantification of gene expression, the in situ localization of gene expression in tissue, and the cloning of genes, as well as for analyzing T-cell clones and the differential expression of genes. For the expert seeking to extend the usefulness of RT-PCR, there are user-friendly applications that complement the latest technological advances, including laser-capture microdissection (LCM), real-time and quantitative PCR, microarray technology, cDNA cloning, and antibody engineering. Study disease pathogenesis with RT-PCR to design new therapeutic strategies Expand RT-PCR with antibody engineering, real-time PCR, and microarray technology.

Some Protocols for CDNA Library Construction with Conifer Tissues

Author : J. A. Pitel
File Size : 69.32 MB
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Surgical Research

Author : Wiley W. Souba
File Size : 22.36 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 ...