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Preface to the Sixth Edition | |
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The Basic Principles of Gene Cloning and DNA Analysis | |
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Why Gene Cloning and DNA Analysis are Important | |
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The early development of genetics | |
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The advent of gene cloning and the polymerase chain reaction | |
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What is gene cloning? | |
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What is PCR? | |
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Why gene cloning and PCR are so important | |
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How to find your way through this book | |
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Vectors for Gene Cloning: Plasmids and Bacteriophages | |
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Plasmids | |
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Bacteriophages | |
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Purification of DNA from Living Cells | |
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Preparation of total cell DNA | |
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Preparation of plasmid DNA | |
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Preparation of bacteriophage DNA | |
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Manipulation of Purified DNA | |
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The range of DNA manipulative enzymes | |
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Enzymes for cutting DNA - restriction endonucleases | |
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Ligation - joining DNA molecules together | |
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Introduction of DNA into Living Cells | |
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Transformation - the uptake of DNA by bacterial cells | |
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Identification of recombinants | |
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Introduction of phage DNA into bacterial cells | |
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Identification of recombinant phages | |
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Introduction of DNA into non-bacterial cells | |
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Cloning Vectors for E. coli | |
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Cloning vectors based on E. coli plasmids | |
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Cloning vectors based on M13 bacteriophage | |
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Cloning vectors based on l bacteriophage | |
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� and other high capacity vectors enable genomic libraries to be constructed | |
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Vectors for other bacteria | |
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Cloning Vectors for Eukaryotes | |
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Vectors for yeast and other fungi | |
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Cloning vectors for higher plants | |
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Cloning vectors for animals | |
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How to Obtain a Clone of a Specific Gene | |
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The problem of selection | |
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Direct selection | |
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Identification of a clone from a gene library | |
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Methods for clone identification | |
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The Polymerase Chain Reaction | |
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The polymerase chain reaction in outline | |
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PCR in more detail | |
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After the PCR: studying PCR products | |
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Real time PCR enables the amount of starting material to be quantified | |
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The Applications of Gene Cloning and DNA Analysis in Research | |
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Sequencing Genes and Genomes | |
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The methodology for DNA sequencing | |
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How to sequence a genome | |
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Studying Gene Expression and Function | |
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Studying the RNA transcript of a gene | |
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Studying the regulation of gene expression | |
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Identifying and studying the translation product of a cloned gene | |
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Studying Genomes | |
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Genome annotation | |
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Studies of the transcriptome and proteome | |
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The Applications of Gene Cloning and DNA Analysis in Biotechnology | |
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Production of Protein from Cloned Genes | |
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Special vectors for expression of foreign genes in E. coli | |
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General problems with the production of recombinant protein in E. coli | |
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Production of recombinant protein by eukaryotic cells | |
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Gene Cloning and DNA Analysis in Medicine | |
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Production of recombinant pharmaceuticals | |
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Identification of genes responsible for human diseases | |
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Gene therapy | |
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Gene Cloning and DNA Analysis in Agriculture | |
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The gene addition approach to plant genetic engineering | |
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Gene subtraction | |
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Problems with genetically modified plants | |
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Gene Cloning and DNA Analysis in Forensic Science and Archaeology | |
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DNA analysis in the identification of crime suspects | |
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Studying kinship by DNA profiling | |
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Sex identification by DNA analysis | |
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Archaeogenetics - using DNA to study human prehistory | |
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Glossary | |
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Index | |