Gene Cloning and DNA Analysis An Introduction

ISBN-10: 1405111216
ISBN-13: 9781405111218
Edition: 5th 2006 (Revised)
Authors: Terry A. Brown
List price: $81.99
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Description: Known world-wide as the standard introductory text to this important and exciting area, the fifth edition of Gene Cloning and DNA Analysis addresses new and growing areas of research whilst retaining the philosophy of the previous editions. Assuming  More...

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Book details

List price: $81.99
Edition: 5th
Copyright year: 2006
Publisher: John Wiley & Sons, Incorporated
Publication date: 3/3/2006
Binding: Paperback
Pages: 408
Size: 7.75" wide x 9.75" long x 1.00" tall
Weight: 2.398
Language: English

Known world-wide as the standard introductory text to this important and exciting area, the fifth edition of Gene Cloning and DNA Analysis addresses new and growing areas of research whilst retaining the philosophy of the previous editions. Assuming the reader has little prior knowledge of the subject its importance, the principles of the techniques used and their applications are all carefully laid out, with over 250 clearly presented two-colour illustrations. In addition to a number of informative changes to the text throughout the book, the final four chapters have been significantly updated and extended to reflect the striking advances made in recent years in the applications of gene cloning and DNA analysis in biotechnology: Extended chapter on agriculture including new material on glyphosate resistant plants New section on the uses of gene cloning and PCR in archaeology Coverage of ethical concerns relating to pharming, gene therapy and GM crops Gene Cloning and DNA Analysis remains an essential introductory text to a wide range of biological sciences students; including genetics and genomics, molecular biology, biochemistry, immunology and applied biology. It is also a perfect introductory text for any professional needing to learn the basics of the subject. All libraries in universities where medical, life and biological sciences are studied and taught should have copies available on their shelves. View the Gene Cloning and DNA Analysis webpage at www.blackwellpublishing.com/genecloning

The Basic Principles of Gene Cloning and Dna Analysis
Why Gene Cloning and DNA Analysis are Important
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
Gene isolation by cloning
Gene isolation by PCR
How to find your way through this book
Vectors for Gene Cloning: Plasmids and Bacteriophages
Plasmids
Basic features of plasmids
Size and copy number
Conjugation and compatibility
Plasmid classification
Plasmids in organisms other than bacteria
Bacteriophages
Basic features of bacteriophages
Lysogenic phages
Gene organization in l DNA molecule
The linear and circular forms of l DNA M13 - a filamentous phage
The attraction of M13 as a cloning vector
Viruses as cloning vectors for other organisms
Purification of DNA from Living Cells
Preparation of total cell DNA
Growing and harvesting a bacterial culture
Preparation of a cell extract
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
Concentration of DNA samples
Measurement of DNA concentration
Other methods for the preparation of total cell DNA
Preparation of plasmid DNA
Separation on the basis of size
Separation on the basis of conformation
Alkaline denaturation
Ethidium bromide-caesium chloride density gradient centrifugation
Plasmid amplification
Preparation of bacteriophage DNA
Growth of cultures to obtain a high l titre
Preparation of non-lysogenic l phages
Collection of phages from an infected culture
Purification of DNA from l phage particles
Purification of M13 DNA causes few problems
Manipulation of Purified DNA
The range of DNA manipulative enzymes
Nucleases
Ligases
Polymerases
DNA modifying enzymes
Topoisomerases
Enzymes for cutting DNA - restriction endonucleases
The discovery and function of restriction endonucleases
Type II restriction endonucleases cut DNA at specific nucleotide sequences
Blunt ends and sticky ends
The frequency of recognition sequences in a DNA molecule
Performing a restriction digest in the laboratory
Analysing the result of restriction endonuclease cleavage
Separation of molecules by gel electrophoresis
Visualizing DNA molecules by staining a gel
Visualizing DNA molecules by autoradiography
Estimation of the sizes of DNA molecules
Mapping the positions of different restriction sites in a DNA molecule
Ligation - joining DNA molecules together
The mode of action of DNA ligase
Sticky ends increase the efficiency of ligation
Putting sticky ends onto a blunt-ended molecule
Linkers
Adaptors
Introduction of DNA into Living Cells
Transformation - the uptake of DNA by bacterial cells
Not all species of bacteria are equally efficient at DNA uptake
Preparation of competent E. coli cells
Selection for transformed cells
Identification of recombinants
Recombinant selection with pBR322 - insertional inactivation of an antibiotic.resistance gene
Insertional inactivation does not always involve antibiotic resistance
Introduction of phage DNA into bacterial cells
Transfection
In vitro packaging of l cloning vectors
Phage infection is visualized as plaques on an agar medium
Identification of recombinant phages
Insertional

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