cDNA Libraries

The numerous vital applications of complementary DNA (cDNA) technology have changed dramatically as the technology has advanced over recent years.Â In cDNA Libraries: Methods and Protocols, expert researchers provide current techniques that reflect the latest advances in the construction and application of cDNA libraries.Â 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.Â Written in the highly successful Methods in Molecular Biology'„¢ 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.Â 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.

Preface	p. v
Contributors	p. ix
Enhanced Approaches to Inherent Problems
Isolation of Fungal Infection Structures from Plant Tissue by Flow Cytometry for Cell-Specific Transcriptome Analysis	p. 3
Preparation of a High-Quality cDNA Library from a Single-Cell Quantity of mRNA Using Chum-RNA	p. 15
Construction of a Full-Length cDNA Library from Castor Endosperm for High-Throughput Functional Screening	p. 37
Full-Length Transcriptome Analysis Using a Bias-Free cDNA Library Prepared with the Vector-Capping Method	p. 53
Construction of Improved Yeast Two-Hybrid Libraries	p. 71
Normalization of Full-Length-Enriched cDNA	p. 85
Bioinformatic Methods for Finding Differentially Expressed Genes in cDNA Libraries, Applied to the Identification of Tumour Vascular Targets	p. 99
Visionary Applications
Enzymatic Production of RNAi Libraries from cDNAs and High-Throughput Selection of Effective shRNA Expression Constructs	p. 123
Construction of Small RNA cDNA Libraries for High-Throughput Sequencing	p. 141
Focusing Mutations Within Random Libraries to Distinct Areas: Protein Domain Library Generation by Overlap Extension	p. 153
Generation of Families of Construct Variants Using Golden Gate Shuffling	p. 167
Application of Full-Length cDNA Resources to Gain-of-Function Technology for Characterization of Plant Gene Function	p. 183
Construction of Yeast Surface-Displayed cDNA Libraries	p. 199
Identification of Protein/Target Molecule Interactions Using Yeast Surface-Displayed cDNA Libraries	p. 211
SNP Discovery by Transcriptome Pyrosequencing	p. 225
RNA-Seq Analysis of Gene Expression and Alternative Splicing by Double-Random Priming Strategy	p. 247
Generation of a Large Catalog of Unique Transcripts for Whole-Genome Expression Analysis in Nonmodel Species	p. 257
Index	p. 271
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