Overview

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.

Full Product Details

Author:   Ian G. Cowell ,  Caroline A. Austin
Publisher:   Humana Press Inc.
Imprint:   Humana Press Inc.
Edition:   1997 ed.
Volume:   69
Dimensions:   Width: 15.50cm , Height: 1.90cm , Length: 23.50cm
Weight:   1.430kg
ISBN:  

9780896033832

ISBN 10:   089603383
Pages:   321
Publication Date:   01 October 1996
Audience:   College/higher education ,  Professional and scholarly ,  Undergraduate ,  Professional & Vocational
Format:   Hardback
Publisher's Status:   Active
Availability:   In Print  
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Table of Contents

cDNA Library Construction from Small Amounts of RNA Using Paramagnetic Beads and PCR.- Increasing the Average Abundance of Low-Abundance cDNAs by Ordered Subdivision of cDNA Populations.- Isolation of Messenger RNA from Plant Tissues.- cDNA Library Construction for the Lambda ZAP®-Based Vectors.- Clone Excision Methods for the Lambda ZAP®-Based Vectors.- Using Rapid Amplification of cDNA Ends (RACE) to Obtain Full-Length cDNAs.- Inverse PCR Approach to Cloning cDNA Ends.- Cloning Gene Family Members Using the Polymerase Chain Reaction with Degenerate Oligonucleotide Primers.- Subtractive Hybridization for the Isolation of Differentially Expressed Genes Using Magnetic Beads.- Preparation of Competent Cells for High-Efficiency Plasmid Transformation of Escherichia coli.- The Tetramethylammonium Chloride (TMAC) Method for Screening cDNA Libraries with Highly Degenerate Oligonucleotide Probes Obtained by Reverse Translation of Amino Acid Sequences.- Screening cDNA Libraries by Hybridization with Double-Stranded DNA Probes and Oligonucleotides.- Immunological Screening of ?-Phage cDNA Expression Libraries.- Cloning Sequence-Specific DNA-Binding Factors from cDNA Expression Libraries Using Oligonucleotide Binding Site Probes.- Protein Interaction Cloning by Far-Western Screening of ?-Phage cDNA Expression Libraries.- Yeast Two-Hybrid Library Screening.- Signal Sequence Trap.- Isolation of Genetic Suppressor Elements (GSEs) from Random Fragment cDNA Libraries in Retroviral Vectors.- Expression and Preparation of Fusion Proteins from Recombinant ?gt1.1 Phages.- Computer Analysis of Cloned Sequences.- The TIGR Human cDNA Database.- Searching the dbEST Database.- Reference cDNA Library Facilities Available from European Sources.

Reviews

This is a good overview of the essentials for preparing a cDNA library, for both the novice and the expert...the book is a true laboratory manual for the experimental scientist. It Should represent a useful and up-to-date addition for any lab that, for whatever reason, needs to generate its own libraries. - Hoffman-LaRoche, Inc. their fine layout and ease of use suggest that the lab copy won't stay on the bookshelf for long. - SGM Quarterly

This is a good overview of the essentials for preparing a cDNA library, for both the novice and the expert...the book is a true laboratory manual for the experimental scientist. It Should represent a useful and up-to-date addition for any lab that, for whatever reason, needs to generate its own libraries.-Hoffman-LaRoche, Inc. ...their fine layout and ease of use suggest that the lab copy won't stay on the bookshelf for long. -SGM Quarterly

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