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cDNA Library Protocols
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Main description:

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.


Contents:

cDNA Library Construction from Small Amounts of RNA Using Paramagnetic Beads and PCR
Kris N. Lambert and Valerie M. Williamson

Increasing the Average Abundance of Low-Abundance cDNAs by Ordered Subdivision of cDNA Populations
David R. Sibson and Michael P. Starkey

Isolation of Messenger RNA from Plant Tissues
Alison Dunn

cDNA Library Construction for the Lambda ZAP (R)-Based Vectors
Marjory A. Snead, Michelle A. Alting-Mees, and Jay M. Short

Clone Excision Methods for the Lambda ZAP (R)-Based Vectors
Marjory A. Snead, Michelle A. Alting-Mees, and Jay M. Short

Using Rapid Amplification of cDNA Ends (RACE) to Obtain Full-Length cDNAs
Yue Zhang and Michael A. Frohman

Inverse PCR Approach to Cloning cDNA Ends
Sheng-He Huang

Cloning Gene Family Members Using the Polymerase Chain Reaction with Degenerate Oligonucleotide Primers
Gregory M. Preston

Subtractive Hybridization for the Isolation of Differentially Expressed Genes Using Magnetic Beads
Hans-Christian Aasheim, Ton Logtenberg, and Frank Larsen

Preparation of Competent Cells for High-Efficiency Plasmid Transformation of Escherichia coli
Yasuhiko Nakata, Xiaoren Tang, and Kazunari K. Yokoyama

The Tetramethylammonium Chloride (TMAC) Method for Screening cDNA Libraries with Highly Degenerate Oligonucleotide Probes Obtained by Reverse Translation of Amino Acid Sequences
Bent Honore and Peder Madsen

Screening cDNA Libraries by Hybridization with Double-Stranded DNA Probes and Oligonucleotides
Caroline A. Austin

Immunological Screening of l-Phage cDNA Expression Libraries
Helen C. Hurst

Cloning Sequence-Specific DNA-Binding Factors from cDNA Expression Libraries Using Oligonucleotide Binding Site Probes
Ian G. Cowell

Protein Interaction Cloning by Far-Western Screening of l-Phage cDNA ExpressionLibraries
Shinichi Takayama and John C. Reed

Yeast Two-Hybrid Library Screening
Ian G. Cowell

Signal Sequence Trap: Expression Cloning Method for Secreted Proteins and Type 1 Membrane Proteins
Kei Tashiro, Toru Nakano, and Tasuku Honjo

Isolation of Genetic Suppressor Elements (GSEs) from Random Fragment cDNA Libraries in Retroviral Vectors
Andrei V. Gudkov and Igor B. Roninson

Expression and Preparation of Fusion Proteins from Recombinant lgt11 Phages
Sheng-He Huang and Ambrose Jong

Computer Analysis of Cloned Sequences
Paul R. Caron

The TIGR Human cDNA Database
Ewen F. Kirkness and Anthony R. Kerlavage

Searching the dbEST Database
Patricia Rodriguez Tome

Reference cDNA Library Facilities Available from European Sources
Michael P. Starkey, Yagnesh Umrania, Christopher R. Mundy, and Martin J. Bishop

Index


PRODUCT DETAILS

ISBN-13: 9781489940506
Publisher: Springer (Humana Press Inc.)
Publication date: August, 2013
Pages: 321
Weight: 487g
Availability: Available
Subcategories: Immunology
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