Preface to the Seventh Edition About the Companion Website Part Ⅰ The Basic Principles of Gene Cloning and DNA Analysis 1 Why Gene Cloning and DNA Analysis are Important 1.1 The early development of genetics 1.2 The advent of gene cloning and the polymerase chain reaction 1.3 What is gene cloning? 1.4 What is PCR? 1.5 Why gene cloning and PCR are so important 1.5.1 Obtaining a pure sample of a gene by cloning 1.5.2 PCR can also be used to purify a gene 1.6 How to find your way through this book Further reading 12 2 Vectors for Gene Cloning: Plasmids and Bacteriophages 2.1 Plasmids 2.1.1 Size and copy number 2.1.2 Conjugation and compatibility 2.1.3 Plasmid classification 2.1.4 Plasmids in organisms other than bacteria 2.2 Bacteriophages 2.2.1 The phage infection cycle 2.2.2 Lysogenic phages Gene organization in the □ DNA molecule The linear and circular forms of □ DNA M13-a filamentous phage 2.2.3 Viruses as cloning vectors for other organisms Further reading 3 Purification of DNA from Living Cells 3.1 Preparation of total cell DNA 3.1.1 Growing and harvesting a bacterial culture 3.1.2 Preparation of a cell extract 3.1.3 Purification of DNA from a cell extract Removing contaminants by organic extraction and enzyme digestion Using ion-exchange chromatography to purify DNA from a cell extract Using silica to purify DNA from a cell extract 3.1.4 Concentration of DNA samples 3.1.5 Measurement of DNA concentration 3.1.6 Other methods for the preparation of total cell DNA 3.2 Preparation of plasmid DNA 3.2.1 Separation on the basis of size 3.2.2 Separation on the basis of conformation Alkaline denaturation Ethidium bromide-caesium chloride density gradient centrifugation 3.2.3 Plasmid amplification 3.3 Preparation of bacteriophage DNA 3.3.1 Growth of cultures to obtain a high □ titre 3.3.2 Preparation of non-lysogenic □ phages 3.3.3 Collection of phages from an infected culture 3.3.4 Purification of DNA from □ phage particles 3.3.5 Purification of M13 DNA causes few problems
Further reading 4 Manipulation of Purified DNA 4.1 The range of DNA manipulative enzymes 4.1.1 Nucleases 4.1.2 Ligases 4.1.3 Polymerases 4.1.4 DNA-modifying enzymes 4.2 Enzymes for cutting DNA: Restriction endonucleases 4.2.1 The discovery and function of restriction endonucleases 4.2.2 Type II restriction endonucleases cut DNA at specific nucleotide sequences 4.2.3 Blunt ends and sticky ends 4.2.4 The frequency of recognition sequences in a DNA molecule 4.2.5 Performing a restriction digest in the laboratory 4.2.6 Analysing the result of restriction endonuclease cleavage Separation of molecules by gel electrophoresis Visualizing DNA molecules in an agarose gel 4.2.7 Estimation of the sizes of DNA molecules 4.2.8 Mapping the positions of different restriction sites in a DNA molecule Part Ⅱ The Applications of Gene Cloning and DNA Analysis in Research Part Ⅲ The Applications of Gene Cloning and DNA Analysis in Biotechnology Glossary Index
[