[...]... Principles of Gene Cloning and DNA Analysis 1 | Why Gene Cloning and DNA Analysis are Important 3 2 | Vectors for Gene Cloning: Plasmids and Bacteriophages 3 | Purification of DNA from Living Cells 25 4 | Manipulation of Purified DNA 45 5 | Introduction of DNA into Living Cells 72 6 | Cloning Vectors for E coli 88 7 | Cloning Vectors for Eukaryotes 105 8 | How to Obtain a Clone of a Specific Gene 126 9 |... nature of the gene until the 1940s Indeed, it was not until Gene Cloning and DNA Analysis: An Introduction 6th edition By T.A Brown Published 2010 by Blackwell Publishing 3 4 Part I The Basic Principles of Gene Cloning and DNA Analysis the experiments of Avery, MacLeod, and McCarty in 1944, and of Hershey and Chase in 1952, that anyone believed that deoxyribonucleic acid (DNA) is the genetic material:... of genes responsible for human diseases 255 14.2.1 How to identify a gene for a genetic disease 256 Locating the approximate position of the gene in the human genome 256 Identification of candidates for the disease gene 258 14.3 Gene therapy 259 14.3.1 Gene therapy for inherited diseases 259 14.3.2 Gene therapy and cancer 260 14.3.3 The ethical issues raised by gene therapy 262 15 Gene Cloning and DNA. .. of a Specific Gene 126 9 | The Polymerase Chain Reaction 147 13 Chapter 1 Why Gene Cloning and DNA Analysis are Important Chapter contents CHAPTER CONTENTS 1.1 1.2 1.3 1.4 1.5 1.6 The early development of genetics The advent of gene cloning and the polymerase chain reaction What is gene cloning? What is PCR? Why gene cloning and PCR are so important How to find you way through this book In the middle... Applications of Gene Cloning and DNA Analysis in Research 163 10 Sequencing Genes and Genomes 165 10.1 The methodology for DNA sequencing 165 10.1.1 Chain termination DNA sequencing 166 Chain termination sequencing in outline 166 Not all DNA polymerases can be used for sequencing 168 Chain termination sequencing requires a single-stranded DNA template 169 The primer determines the region of the template DNA that... sensitivity means that PCR can also be used with DNA from forensic material such as hairs and dried bloodstains or even from the bones of long-dead humans (Chapter 16) Chapter 1 Why Gene Cloning and DNA Analysis are Important 1.6 How to find your way through this book This book explains how gene cloning, PCR and other DNA analysis techniques are carried out and describes the applications of these techniques... (e.g M13) 17 Part I The Basic Principles of Gene Cloning and DNA Analysis 18 Phage particle DNA 1 The phage attaches to the bacterium and injects its DNA Phage DNA molecule 2 The phage DNA molecule is replicated 3 Capsid components are synthesized, new phage particles are assembled and released Capsid components Cell lysis New phage particle Figure 2.6 The general pattern of infection of a bacterial... evolution and the impact of environmental change on the biosphere, and to bring their powerful tools to bear in the fight against crime Forty years have passed since the dawning of the age of gene cloning, but we are still riding the rollercoaster and there is no end to the excitement in sight Chapter 1 Why Gene Cloning and DNA Analysis are Important Figure 1.1 Construction of a recombinant DNA molecule... double-stranded DNA molecules, each of which consists of one strand of the original molecule and one new strand of DNA, denature into single strands This begins a second cycle of denaturation–annealing–synthesis, at the end of which there are eight DNA strands By repeating the cycle 30 times the double-stranded molecule that we began with is converted into over 130 million new double-stranded molecules,... 1.5 Why gene cloning and PCR are so important As you can see from Figures 1.1 and 1.2, gene cloning and PCR are relatively straightforward procedures Why, then, have they assumed such importance in biology? The answer is largely because both techniques can provide a pure sample of an individual gene, separated from all the other genes in the cell 1.5.1 Obtaining a pure sample of a gene by cloning To . from Cloned Genes 225 14 Gene Cloning and DNA Analysis in Medicine 245 15 Gene Cloning and DNA Analysis in Agriculture 264 16 Gene Cloning and DNA Analysis. 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