PCR Cloning Protocols, 2nd ed.

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publication info

B.Y. Chan and H.W. Janes, ed., Humana Press, 2002, 421 pp., comb bound

mfr. no.



PCR: suitable

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General description

This edition update and expands Bruce White′s best-selling "PCR Protocols" (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long PCR and GC-rich template amplification. Powerful applications of PCR in library construction and sublibrary generation and screening are presented.

Table of Contents

Part I. Performing and Optimizing PCR
Polymerase Chain Reaction: Basic Principles and Routine Practice
Computer Programs for PCR Primer Design and Analysis
Single-Step PCR Optimization Using Touchdown and Stepdown PCR Programming
XL PCR Amplification of Long Targets from Genomic DNA
Coupled One-Step Reverse Transcription and Polymerase Chain Reaction Procedure for Cloning Large cDNA Fragments
Long Distance Reverse-Transcription PCR
Increasing PCR Sensitivity for Amplification from Paraffin-Embedded Tissues
GC-Rich Template Amplification by Inverse PCR: DNA Polymerase and Solvent Effects
PCR Procedure for the Isolation of Trinucleotide Repeats
Methylation-Specific PCR
Direct Cloning of Full-Length Cell Differentially Expressed Genes by Multiple Rounds of Subtractive Hybridization Based on Long-Distance PCR and Magnetic Beads
Part II. Cloning PCR Products
Cloning PCR Products: An Overview
Using T4 DNA Polymerase to Generate Clonable PCR Products
Enzyme-Free Cloning of PCR Products and Fusion Protein Expression
Directional Restriction Site-Free Insertion of PCR Products into Vectors
Autosticky PCR: Directional Cloning of PCR Products with Preformed 5′ Overhangs
A Rapid and Simple Procedure for Direct Cloning of PCR Products into Baculoviruses
Part III. Mutagenesis and Recombination
PCR Approaches to DNA Mutagenesis and Recombination: An Overview
In-Frame Cloning of Synthetic Genes Using PCR Inserts
Megaprimer PCR PCR-Mediated Recombination: A General Method Applied to Construct Chimeric Infectious Molecular Clones
PCR Method for Generating Multiple Mutations at Adjacent Sites
A Fast Polymerase Chain Reaction-Mediated Strategy for Introducing Repeat Expansions into CAG-Repeat Containing Genes
PCR Screening in Signature-Tagged Mutagenesis of Essential Genes
Staggered Extension Process (StEP) In Vitro Recombination
Random Mutagenesis by Whole-Plasmid PCR Amplification
Part IV. Cloning Unknown Neighboring DNA
PCR-Based Strategies to Clone Unknown DNA Regions from Known Foreign Integrants: An Overview
Long Distance Vectorette PCR (LDV PCR) Nonspecific, Nested Suppression PCR Method for Isolation of Unknown Flanking DNA ("Cold-Start Method")
Inverse PCR: cDNA Cloning
Inverse PCR: Genomic DNA Cloning
Gene Cloning and Expression Profiling by Rapid Amplification of Gene Inserts with Universal Vector Primers
The Isolation of DNA Sequences Flanking Tn5 Transposon Insertions by Inverse PCR
Rapid Amplification of Genomic DNA Sequences Tagged by Insertional Mutagenesis
Isolation of Large Terminal Sequences of BAC Inserts Based on Double-Restriction-Enzyme Digestion Followed by Anchored PCR
A "Step Down" PCR-Based Technique for Walking Into and the Subsequent Direct Sequence Analysis of Flanking Genomic DNA
Part V. Library Construction and Screening
Use of PCR in Library Screening: An Overview
Cloning of Homologous Genes by Gene-Capture PCR
Rapid and Nonradioactive Screening of Recombinant Libraries by PCR
Rapid cDNA Cloning by PCR Screening (RC-PCR) Generation and PCR Screening of Bacteriophage l Sublibraries Enriched for Rare Clones (the "Sublibrary Method") PCR-Based Screening for Bacterial Artificial Chromosome Libraries
A 384-Well Microtiter-Plate-Based Template Preparation and Sequencing Method
A Microtiter-Plate-Based High Throughput PCR Product Purification Method
Certificate of Analysis
Certificate of Origin

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