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Known worldwide as the standard introductory text to this important and exciting area of study, Gene Cloning and DNA Analysis: An Introduction, 8th Edition preserves the tradition of excellence created by previous editions. Comprehensive and authoritative, the book explores all of the topics crucial to an understanding of gene cloning in an approachable way. An easy-to-follow and user-friendly layout is presented in full-color throughout the volume, making it simple to absorb the clear and accessible material contained within.
Gene Cloning and DNA Analysis: An Introduction, 8th Edition contains updated and extended coverage of gene editing strategies like CRISPR/Cas, rewritten chapters on DNA sequencing and genome studies, as well as new material on real-time PCR and typing of human disease mutations. Over 250 full-color illustrations are included to bring to life the comprehensive content. The book also covers topics like:
The strategies used by researchers and industry practitioners to assemble genome sequences
Next generation sequencing methods and descriptions of their applications in studying genomes and transcriptomes
Includes the use and application of gene editing strategies
Interbreeding between Neanderthals and Homo Sapiens
Gene Cloning and DNA Analysis: An Introduction, 8th Edition is an invaluable introductory text for students in classes like genetics and genomics, molecular biology, biochemistry, immunology, and applied biology. It also belongs on the bookshelves of every professional who desires to improve their understanding of the basics of gene cloning or DNA analysis.
Autorentext
T. A. Brown is Emeritus Professor of Biomolecular Archaeology in the Manchester Institute of Biotechnology at the University of Manchester in the United Kingdom. He has published several books on genetics, genomics and biochemistry as well as over 150 research papers.
Klappentext
Known worldwide as the standard introductory text to cloning and DNA analysis in Genetics The eighth edition of this well-recognised textbook preserves the tradition of excellence created by previous editions. Comprehensive and authoritative, the book explores all of the topics crucial to an understanding of gene cloning in an approachable way. An easy-to-follow and user-friendly layout is presented in full-colour throughout the volume, making it simple to absorb the clear and accessible material contained within. Gene Cloning and DNA Analysis: An Introduction, Eighth Edition contains updated and extended coverage of gene editing strategies like CRISPR/Cas, rewritten chapters on DNA sequencing and genome studies, as well as new material on real-time PCR and typing of human disease mutations. Over 250 full-colour illustrations are included to bring to life the comprehensive content. The book also covers topics like:
Inhalt
Preface to the Eighth Edition xv
Part I The Basic Principles of Gene Cloning and DNA Analysis 1
1 Why Gene Cloning and DNA Analysis are Important 3
2 Vectors for Gene Cloning: Plasmids and Bacteriophages 15
3 Purification of DNA from Living Cells 29
4 Manipulation of Purified DNA 53
5 Introduction of DNA into Living Cells 83
6 Cloning Vectors for E. coli 101
7 Cloning Vectors for Eukaryotes 121
8 How to Obtain a Clone of a Specific Gene 145
9 The Polymerase Chain Reaction 169
Part II The Applications of Gene Cloning and DNA Analysis in Research 187
10 Sequencing Genes and Genomes 189
11 Studying Gene Expression and Function 217
12 Studying Genomes 243
13 Studying Transcriptomes and Proteomes 259
Part III The Applications of Gene Cloning and DNA Analysis in Biotechnology 275
14 Production of Protein from Cloned Genes 277
15 Gene Cloning and DNA Analysis in Medicine 301
16 Gene Cloning and DNA Analysis in Agriculture 327
17 Gene Cloning and DNA Analysis in Forensic Science and Archaeology 355
Glossary 377
Index 395
Preface to the Eighth Edition xv
Part I The Basic Principles of Gene Cloning and DNA Analysis 1
1 Why Gene Cloning and DNA Analysis are Important 3
1.1 The early development of genetics 4
1.2 The advent of gene cloning and the polymerase chain reaction 4
1.3 What is gene cloning? 5
1.4 What is PCR? 5
1.5 Why gene cloning and PCR are so important 8
1.5.1 Obtaining a pure sample of a gene by cloning 8
1.5.2 PCR can also be used to purify a gene 10
1.6 How to find your way through this book 11
Further reading 13
2 Vectors for Gene Cloning: Plasmids and Bacteriophages 15
2.1 Plasmids 15
2.1.1 Size and copy number 17
2.1.2 Conjugation and compatibility 18
2.1.3 Plasmid classification 19
2.1.4 Plasmids in organisms other than bacteria 19
2.2 Bacteriophages 19
2.2.1 The phage infection cycle 20
2.2.2 Lysogenic phages 20
2.2.3 Viruses as cloning vectors for other organisms 26
Further reading 27
3 Purification of DNA from Living Cells 29
3.1 Preparation of total cell DNA 30
3.1.1 Growing and harvesting a bacterial culture 30
3.1.2 Preparation of a cell extract 31
3.1.3 Purification of DNA from a cell extract 33
3.1.4 Concentration of DNA samples 37
3.1.5 Measurement of DNA concentration 38
3.1.6 Other methods for the preparation of total cell DNA 39
3.2 Preparation of plasmid DNA 40
3.2.1 Separation on the basis of size 41
3.2.2 Separation on the basis of conformation 42
3.2.3 Plasmid amplification 44
3.3 Preparation of bacteriophage DNA 46
3.3.1 Growth of cultures to obtain a high titre 47
3.3.2 Preparation of nonlysogenic phages 47
3.3.3 Collection of phages from an infected culture 49
3.3.4 Purification of DNA from phage particles 49
3.3.5 Purification of M13 DNA causes few problems 49
Further reading 51
4 Manipulation of Purified DNA 53
4.1 The range of DNA manipulative enzymes 55
4.1.1 Nucleases 55
4.1.2 Ligases 57
4.1.3 Polymerases 57
4.1.4 DNA modifying enzymes 58
4.2 Enzymes for cutting DNA restriction endonucleases 59
4.2.1 The discovery and function of restriction endonucleases 60
4.2.2 Type II restriction endonucleases cut DNA at specific nucleotide sequences 61
4.2.3 Blunt ends and sticky ends 62
4.2.4 The frequency of recognition sequences in a DNA molecule 63 4.2.5 ...