Genetic Library Construction and Screening

by Bird, R. Curtis; Smith, Bruce F.

• ISBN13: 9783540672784
• ISBN10: 3540672788
• Edition: 1st
• Format: Paperback
• Copyright: 2002-07-01
• Publisher: Springer Nature

Summary

This volume is designed to provide an easily accessible introduction to 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, such as variations on two-hybrid analysis for investigation of protein-protein interactions or investigation of protein-nucleic acid interactions.

Table of Contents

PCR and cDNA Cloning Techniques
Strategies for cDNA Cling and Mapping RNA Transcripts	p. 3
Cloning PCR Products	p. 21
Site Directed Deletion,Insertion, and Substitution Using PCR	p. 33
Long RT-PCR Cloning - Amplification of Full-Length Enterovirus Genomes	p. 55
Construction of cDNA Libraries from Small Quantities of Total RNA Using Template Switching Catalyzed by M-MLV Reverse Transcriptase	p. 69
Subtractive and Differential Display Techniques
Subtractive Hybridization and cDNA Cloning	p. 97
Differential Display to Identify Steroid-Induced Genes in Endocrine Cells	p. 127
Specialized Cloning and Library Screening Strategies
Two Hyrid cDNA Cloning	p. 145
Yeast One and Two Hyrid cDNA Cloning	p. 167
High-Throughput Library Screening by Fluorescent Hybridisations on Gridded Membranes	p. 187
Identification of Cell Targeting Ligands Using Random Peptide-Presenting Phage Libraries	p. 209
Subject Index	p. 233
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