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Informationen zum Autor Jeremy W. Dale is a professor emeritus in the Microbial and Cellular Sciences Department at the University of Surrey, UK. Malcolm von Schantz is Professor of Chronobiology at the University of Surrey. He is an internationally recognised researcher and an experienced educator, who received his training in Sweden, the United States, and the UK. Nicholas Plant is the author of From Genes to Genomes: Concepts and Applications of DNA Technology , 3rd Edition, published by Wiley. Klappentext The latest edition of this highly successful textbook introduces the key techniques and concepts involved in cloning genes and in studying their expression and variation. The new edition features: Increased coverage of whole-genome sequencing technologies and enhanced treatment of bioinformatics. Clear, two-colour diagrams throughout. A dedicated website including all figures. Noted for its outstanding balance between clarity of coverage and level of detail, this book provides an excellent introduction to the fast moving world of molecular genetics. Zusammenfassung The latest edition of this highly successful textbook introduces the key techniques and concepts involved in cloning genes and in studying their expression and variation. The new edition features: * Increased coverage of whole-genome sequencing technologies and enhanced treatment of bioinformatics. Inhaltsverzeichnis Preface xiii 1 From Genes to Genomes 1 1.1 Introduction 1 1.2 Basic molecular biology 4 1.2.1 The DNA backbone 4 1.2.2 The base pairs 6 1.2.3 RNA structure 10 1.2.4 Nucleic acid synthesis 11 1.2.5 Coiling and supercoilin 11 1.3 What is a gene? 13 1.4 Information flow: gene expression 15 1.4.1 Transcription 16 1.4.2 Translation 19 1.5 Gene structure and organisation 20 1.5.1 Operons 20 1.5.2 Exons and introns 21 1.6 Refinements of the model 22 2 How to Clone a Gene 25 2.1 What is cloning? 25 2.2 Overview of the procedures 26 2.3 Extraction and purification of nucleic acids 29 2.3.1 Breaking up cells and tissues 29 2.3.2 Alkaline denaturation 31 2.3.3 Column purification 31 2.4 Detection and quantitation of nucleic acids 32 2.5 Gel electrophoresis 33 2.5.1 Analytical gel electrophoresis 33 2.5.2 Preparative gel electrophoresis 36 2.6 Restriction endonucleases 36 2.6.1 Specificity 37 2.6.2 Sticky and blunt ends 40 2.7 Ligation 42 2.7.1 Optimising ligation conditions 44 2.7.2 Preventing unwanted ligation: alkaline phosphatase and double digests 46 2.7.3 Other ways of joining DNA fragments 48 2.8 Modification of restriction fragment ends 49 2.8.1 Linkers and adaptors 50 2.8.2 Homopolymer tailing 52 2.9 Plasmid vectors 53 2.9.1 Plasmid replication 54 2.9.2 Cloning sites 55 2.9.3 Selectable markers 57 2.9.4 Insertional inactivation 58 2.9.5 Transformation 59 2.10 Vectors based on the lambda bacteriophage 61 2.10.1 Lambda biology 61 2.10.2 In vitro packaging 65 2.10.3 Insertion vectors 66 2.10.4 Replacement vectors 68 2.11 Cosmids 71 2.12 Supervectors: YACs and BACs 72 2.13 Summary 73 3 Genomic and cDNA Libraries 75 3.1 Genomic libraries 77 3.1.1 Partial digests 77 3.1.2 Choice of vectors 80 3.1.3 Construction and evaluation of a genomic library 83 3.2 Growing and storing libraries 86 3.3 cDNA libraries 87 3.3.1 Isolation of mRNA 88 3.3.2 cDNA synthesis 89 3.3.3 Bacterial cDNA 93 3.4 Screening libraries with gene probes 94 3.4.1 Hybridization 94 ...
