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, cDNA Library Protocols offers a wide range of approaches to screening cDNA libraries.
- 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 Expression Libraries; 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
"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