Complementary DNA Libraries -- Rapid Amplification of cDNA Ends -- cDNA Generation on Paramagnetic Beads -- Construction of a Normalized cDNA Library by mRN-cDNA Hybridization and Subtraction -- Amplification of cDNA Ends Using PCR Suppression Effect and Step-Out PCR -- Use of Inverse PCR to Clone cDNA Ends -- Construction of Size-Fractionated cDNA Library Assisted by an In Vitro Recombination Reaction -- Construction of a Full-Length Enriched and a 5?-End Enriched cDNA Library Using the Oligo-Capping Method -- cDNA Library Construction Using In Vitro Transcriptional Amplification -- Amplification of Representative cDNA Pools from Microscopic Amounts of Animal Tissue -- Single-Cell cDNA Library Construction Using Cycling aRNA Amplification -- mRNA/cDNA Library Construction Using RNA-Polymerase Cycling Reaction -- Quality Assessment of cDNA Libraries -- Assessment of the Quality of mRNA Libraries by Agarose Gel Electrophoresis -- PACS RT-PCR -- Single-Cell mRNA Library Analysis by Northern Blot Hybridization -- Generation of cDNA Libraries for Profiling Gene Expression of Given Tissues or Cells -- Screening Poly [dA/dT(?)] cDNA for Gene Identification -- Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification -- Generation of Full-Length cDNA Libraries Enriched for Differentially Expressed Genes -- Subtractive Hybridization for the Identification of Differentially Expressed Genes Using Uraci-DNA Glycosylase and Mung-Bean Nuclease -- Subtractive Cloning of Differential Genes Using RNA-PCR -- Strategy for Construction of a cDNA Encoding a Repetitive Amino Acid Sequence -- Preparing Lambda Libraries for Expression of Proteins in Prokaryotes or Eukaryotes -- Peptide Library Construction from RNA-PCR-Derived RNAs -- Identifying Interacting Proteins in an Escherichia coli-Based Two-Hybrid System -- Future Perspectives.

With the decoding of the human genome and the potential application of its information to gene chips, the ability to generate cDNA libraries for functional assays of specific gene sequences has become critical in gene manipulation and genetic research. In Generation of cDNA Libraries: Methods and Protocols, expert researchers and inventors in the field describe their own proven techniques for generating cDNA/mRNA libraries to identify the functions of specific decoded gene sequences. A wide variety of techniques is presented for enhancing the generation of complete and full-length libraries, and for confirming the quality of the cDNAs generated. Among the applications detailed are electrophoresis, Northern blotting, single-cell microarray analysis, subtractive hybridization, subtractive cloning, gene cloning, and peptide library generation. Each method includes background information, step-by-step instructions, a list of reagents, operational tips, and notes on instrumentation. A background tutorial provides essential information: the definition of a cDNA library, the various types of cDNA libraries, the differences between methods for cDNA library generation using either conventional approaches or novel strategies, and the quality of cDNA libraries. State-of-the-art and highly practical, Generation of cDNA Libraries: Methods and Protocols provides a variety of powerful and readily reproducible techniques for the generation of the entire range of complete, full-length cDNA/mRNA libraries needed in today's forefront genetic research.