Cdna Library Protocols

Front Cover
Ian G. Cowell
Springer Science & Business Media, 1997 - Antisense DNA - 321 pages
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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, the book offers a wide range of approaches to screening cDNA libraries.
 

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Contents

cDNA Library Construction from Small Amounts of RNA Using Paramagnetic Beads and PCR
1
Increasing the Average Abundance of LowAbundance cDNAs by Ordered Subdivision of cDNA Populations
13
Isolation of Messenger RNA from Plant Tissues
33
cDNA Library Construction for the Lambda ZAPBased Vectors
39
Clone Excision Methods for the Lambda ZAPBased Vectors
53
Using Rapid Amplification of cDNA Ends RACE to Obtain FullLength cDNAs
61
Inverse PCR Approach to Cloning cDNA Ends
89
Cloning Gene Family Members Using the Polymerase Chain Reaction with Degenerate Oligonucleotide Primers
97
Cloning SequenceSpecific DNABinding Factors from cDNA Expression Libraries Using Oligonucleotide Binding Site Probes
161
Protein Interaction Cloning by FarWestern Screening of 𝝀Phage cDNA Expression Libraries
171
Yeast TwoHybrid Library Screening
185
Signal Sequence Trap Expression Cloning Method for Secreted Proteins and Type 1 Membrane Proteins
203
Isolation of Genetic Suppressor Elements GSEs from Random Fragment cDNA Libraries in Retroviral Vectors
221
Expression and Preparation of Fusion Proteins from Recombinant 𝝀gt11 Phages
241
Computer Analysis of Cloned Sequences Paul R Caron
247
The TIGR Human cDNA Database
261

Subtractive Hybridization for the Isolation of Differentially Expressed Genes Using Magnetic Beads
115
Preparation of Competent Cells for HighEfficiency Plasmid Transformation of Escherichia coli
129
The Tetramethylammonium Chloride TMAC Method for Screening cDNA Libraries with Highly Degenerate Oligonucleotide Probes Obtained by Re...
139
Screening cDNA Libraries by Hybridization with DoubleStranded DNA Probes and Oligonucleotides
147
Immunological Screening of 𝝀Phage cDNA Expression Libraries
155
Searching the dbEST Database
269
Reference cDNA Library Facilities Available from European Sources
285
Index
317
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