cDNA Library Protocols
cDNA Library Protocols

Author: Ian G. Cowell

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 324

ISBN-13: 1592595553

DOWNLOAD EBOOK

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.

Generation of cDNA Libraries
Generation of cDNA Libraries

Author: Shao-Yao Ying

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 329

ISBN-13: 1592593593

DOWNLOAD EBOOK

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.

CDNA Library Protocols. Methods in Molecular Biology
CDNA Library Protocols. Methods in Molecular Biology

Author: Ian G. Cowell

Publisher:

Published: 1997

Total Pages:

ISBN-13:

DOWNLOAD EBOOK

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.

cDNA Libraries
cDNA Libraries

Author: Chaofu Lu

Publisher: Humana Press

Published: 2011-03-03

Total Pages: 275

ISBN-13: 9781617790645

DOWNLOAD EBOOK

The numerous vital applications of complementary DNA (cDNA) technology have changed dramatically as the technology has advanced over recent years. In cDNA Libraries: Methods and Protocols, expert researchers provide current techniques that reflect the latest advances in the construction and application of cDNA libraries. The first half of the volume covers improved approaches to some of the most basic elements of creating cDNA libraries, while the second half casts a much wider net and includes visionary applications of cDNA technology which were either unforeseen or technically impractical until recently. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, cDNA Libraries: Methods and Protocols serves as an ideal guide to all scientists seeking to advance this important technology and provide answers to the enduring fundamental questions of biology.

The Nucleic Acid Protocols Handbook
The Nucleic Acid Protocols Handbook

Author: Ralph Rapley

Publisher: Springer Science & Business Media

Published: 2008-06-29

Total Pages: 997

ISBN-13: 1592590381

DOWNLOAD EBOOK

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."

Short Protocols in Molecular Biology
Short Protocols in Molecular Biology

Author: Frederick M. Ausubel

Publisher: Wiley-Interscience

Published: 1989-09-25

Total Pages: 422

ISBN-13:

DOWNLOAD EBOOK

This volume contains shortened versions of the methods published in the looseleaf manual Current Protocols in Molecular Biology. It presents fully-tested, current techniques based on material from the core manual and from the quarterly update service. Includes all step-by-step descriptions of methods covered in the first ten chapters of CPMB and provides enough detail to perform the experiments (only introductions, annotations, and commentary have been omitted). Marginal notes explain the hows and whys of many steps, and provide tips on safety, storage, and anticipated results. Includes references and recipes for all reagents and media and helpful tables and illustrations.

Natural Killer Cell Protocols
Natural Killer Cell Protocols

Author: Kerry S. Campbell

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 394

ISBN-13: 1592590446

DOWNLOAD EBOOK

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.

RT-PCR Protocols
RT-PCR Protocols

Author: Nicola King

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 370

ISBN-13: 159259283X

DOWNLOAD EBOOK

Until the mid 1980s, the detection and quantification of a specific mRNA was a difficult task, usually only undertaken by a skilled molecular biologist. With the advent of PCR, it became possible to amplify specific mRNA, after first converting the mRNA to cDNA via reverse transcriptase. The arrival of this technique—termed reverse transcription-PCR (RT-PCR)—meant that mRNA suddenly became amenable to rapid and sensitive analysis, without the need for advanced training in molecular biology. This new accessibility of mRNA, which has been facilitated by the rapid accumulation of sequence data for human mRNAs, means that every biomedical researcher can now include measurement of specific mRNA expression as a routine component of his/her research plans. In view of the ubiquity of the use of standard RT-PCR, the main objective of RT-PCR Protocols is essentially to provide novel, useful applications of RT-PCR. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the basic RT-PCR protocol. For example, a variety of different adaptations are described that have been employed to obtain quantitative data from RT-PCR. Quantitative RT-PCR provides the ability to accurately measure changes/imb- ances in specific mRNA expression between normal and diseased tissues.