Generation of cDNA Libraries

Generation of cDNA Libraries
Author: Shao-Yao Ying
Publisher: Springer Science & Business Media
Total Pages: 329
Release: 2008-02-03
Genre: Science
ISBN: 1592593593

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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.

cDNA Library Protocols

cDNA Library Protocols
Author: Ian G. Cowell
Publisher: Springer Science & Business Media
Total Pages: 324
Release: 2008-02-02
Genre: Science
ISBN: 1592595553

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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 Libraries

cDNA Libraries
Author: Chaofu Lu
Publisher: Humana Press
Total Pages: 275
Release: 2016-08-23
Genre: Medical
ISBN: 9781493957965

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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.

Biotechnology

Biotechnology
Author: B.N. Pandey
Publisher: APH Publishing
Total Pages: 426
Release: 2008
Genre: Biotechnology
ISBN: 9788131303061

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Genetic Library Construction and Screening

Genetic Library Construction and Screening
Author: R.C. Bird
Publisher: Springer Science & Business Media
Total Pages: 244
Release: 2012-12-06
Genre: Science
ISBN: 3642564089

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Designed as an introductory text the authors cover all core strategies in the application of modern recombinant DNA technology. The first chapters directly address the applications of polymerase chain reaction to a variety of problems in DNA cloning that are, or have been, extremely challenging using more traditional approaches and technologies. These include cDNA cloning and transcript mapping, mutagenesis as well as the cloning of very long transcripts and protocols using limiting amounts of total RNA. Further chapters describe approaches to subtractive cloning technologies as well as novel specialized expression cloning and library screening strategies. The handbook contains detailed step-by-step protocols and extensive hands-on advice.

Advances in Animal Genomics

Advances in Animal Genomics
Author: Sukanta Mondal
Publisher: Academic Press
Total Pages: 340
Release: 2020-11-25
Genre: Science
ISBN: 0128206128

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Advances in Animal Genomics provides an outstanding collection of integrated strategies involving traditional and modern - omics (structural, functional, comparative and epigenomics) approaches and genomics-assisted breeding methods which animal biotechnologists can utilize to dissect and decode the molecular and gene regulatory networks involved in the complex quantitative yield and stress tolerance traits in livestock. Written by international experts on animal genomics, this book explores the recent advances in high-throughput, next-generation whole genome and transcriptome sequencing, array-based genotyping, and modern bioinformatics approaches which have enabled to produce huge genomic and transcriptomic resources globally on a genome-wide scale. This book is an important resource for researchers, students, educators and professionals in agriculture, veterinary and biotechnology sciences that enables them to solve problems regarding sustainable development with the help of current innovative biotechnologies. Integrates basic and advanced concepts of animal biotechnology and presents future developments Describes current high-throughput next-generation whole genome and transcriptome sequencing, array-based genotyping, and modern bioinformatics approaches for sustainable livestock production Illustrates integrated strategies to dissect and decode the molecular and gene regulatory networks involved in complex quantitative yield and stress tolerance traits in livestock Ensures readers will gain a strong grasp of biotechnology for sustainable livestock production with its well-illustrated discussion

Genomics Protocols

Genomics Protocols
Author: Michael P. Starkey
Publisher: Springer Science & Business Media
Total Pages: 537
Release: 2008-02-03
Genre: Medical
ISBN: 159259235X

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We must unashamedly admit that a large part of the motivation for editing Genomics Protocols was selfish. The possibility of assembling in a single volume a unique and comprehensive collection of complete protocols, relevant to our work and the work of our colleagues, was too good an opportunity to miss. We are pleased to report, however, that the outcome is something of use not only to those who are experienced practitioners in the genomics field, but is also valuable to the larger community of researchers who have recognized the potential of genomics research and may themselves be beginning to explore the technologies involved. Some of the techniques described in Genomics Protocols are clearly not restricted to the genomics field; indeed, a prerequisite for many procedures in this discipline is that they require an extremely high throughput, beyond the scope of the average investigator. However, what we have endeavored here to achieve is both to compile a collection of procedures concerned with geno- scale investigations and to incorporate the key components of “bottom-up” and “top-down” approaches to gene finding. The technologies described extend from those traditionally recognized as coming under the genomics umbrella, touch on proteomics (the study of the expressed protein complement of the genome), through to early therapeutic approaches utilizing the potential of genome programs via gene therapy (Chapters 27–30).

The Practical Bioinformatician

The Practical Bioinformatician
Author: Limsoon Wong
Publisher: World Scientific
Total Pages: 539
Release: 2004
Genre: Medical
ISBN: 981238846X

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Computer scientists have increasingly been enlisted as "bioinformaticians" to assist molecular biologists in their research. This book is a practical introduction to bioinformatics for these computer scientists. The chapters are in-depth discussions by expert bioinformaticians on both general techniques and specific approaches to a range of selected bioinformatics problems. The book is organized into clusters of chapters on the following topics: - Overview of modern molecular biology and a broad spectrum of techniques from computer science -- data mining, machine learning, mathematical modeling, sequence alignment, data integration, workflow development, etc. - In-depth discussion of computational recognition of functional and regulatory sites in DNA sequences. - Incisive discussion of computational prediction of secondary structure of RNA sequences. - Overview of computational prediction of protein cellular localization, and selected discussions of inference of protein function. - Overview of methods for discovering protein-protein interactions. - Detailed discussion of approaches to gene expression analysis for the diagnosis of diseases, the treatment of diseases, and the understanding of gene functions. - Case studies on analysis of phylogenies, functional annotation of proteins, construction of purposebuilt integrated biological databases, and development of workflows underlying the large-scale-effort gene discovery. - Written in a practical, in-depth tutorial style - Covers a broad range of bioinformatics topics and of techniques used in bioinformatics - Comprehensive overviews of the development of various approaches in a number of selectedtopics - In-depth exposition of a number of important topics - Contributions by prominent researchers: Vladimir Bajic, Ming Li, Kenta Nakai, Limsoon Wong, Cathy Wu, etc. - Extensive, integrated references to background liter

Next-Generation Sequencing Data Analysis

Next-Generation Sequencing Data Analysis
Author: Xinkun Wang
Publisher: CRC Press
Total Pages: 252
Release: 2016-04-06
Genre: Mathematics
ISBN: 1482217899

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A Practical Guide to the Highly Dynamic Area of Massively Parallel SequencingThe development of genome and transcriptome sequencing technologies has led to a paradigm shift in life science research and disease diagnosis and prevention. Scientists are now able to see how human diseases and phenotypic changes are connected to DNA mutation, polymorphi