Essential Molecular Biology / Edition 2

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Overview


The two Essential Molecular Biology books in the Practical Approach Series are designed for the absolute beginner at gene cloning whether they be at the start of their career or an experienced researcher in another field. As with the first editions, the objective of both volumes is to combine solid practical information with sufficient background material to ensure that the novice can understand how a technique works, what it achieves, and how to make modifications to suit personal requirements. Volume 1 concentrates on the procedures for DNA and RNA manipulation: purification, electrophoresis, and the construction and cloning of recombinant molecules. It also includes a general introduction to molecular biology in the laboratory and a survey of cloning vectors for Escherichia Coli.

The book contains black-and-white illustrations.

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Editorial Reviews

From The Critics
A handbook for the absolute beginner at cloning genes, either because they are just entering molecular biology, or because they work in a related field of biology and their friend who used to do that for them just dropped dead of some mysterious disease. The first volume deals with the fundamental techniques needed to carry out DNA cloning experiments, emphasizing practical skills and enough background to adjust to circumstances as the research project progresses. The second volume builds on the first by describing procedures for preparing gene libraries and identifying genes. The second edition incorporates the expansion in the range and sophistication of molecular biology techniques since 1990. Annotation c. Book News, Inc., Portland, OR
Booknews
Presents a practical manual designed for beginners to gene cloning and recombinant DNA techniques. Practical information is combined with background material to allow readers to understand how techniques work and how to make modifications to suit personal requirements. Concentrates on procedures for DNA and RNA manipulation, including purification, electrophoresis, and the construction and cloning of recombinant molecules. Includes an introduction to molecular biology in the laboratory and a survey of cloning vectors for , recipes and general procedures, information on DNA and RNA modification enzymes, a list of restriction endonucleases, and a list of suppliers. Brown teaches biomolecular sciences at UMIST, UK. Annotation c. Book News, Inc., Portland, OR (booknews.com)
Doody's Review Service
Reviewer: Dean Rosenthal, PhD (Georgetown University School of Medicine)
Description: This is a book for absolute beginners in the field of molecular biology.
Purpose: The book describes basic protocols in molecular biology, including culturing of host strains of bacteria and bacteriophage, purification of DNA and RNA, electrophoresis of nucleic acids, and the generation and isolation recombinant DNA.
Audience: There are numerous tidbits of information throughout the book for the novice and experienced molecular biologist alike, such as the usefulness of shrimp alkaline phosphatase, which is more heat labile than CIP, for cloning.
Features: One of the strengths of the manual is that many different protocols for the same procedure are supplied (e.g. RNA isolation). In addition, the book includes explanations of protocols that are the basis of commercially available molecular biology "kits", such as those sold for RNA isolation, and PCR cloning. While the editor discourages the over-reliance on these kits, mention is made of the level of difficulty of certain protocols that may require their use, including T/A cloning. Protocols are clearly boxed in gray, and all are relatively easy to understand. One minor flaw is its brevity, as well as the sometimes less-than-perfectly integrated nature of a protocol book with contributions by different authors. For example, in some chapters, the authors supply their own ingredients for their protocols; others refer to appendix; while sometimes there is no reference to either. For example, in chapter 2, protocol 1, it states to "Select the ingredients for the required agar medium", without reference to Appendix II in the back of the book. Also, each chapter restarts with protocol 1 again. It would therefore be helpful to at least have the chapter numbers at the header or footer of each page.
Assessment: These minor criticisms aside, this is a concise, practical reference manual for recombinant DNA techniques.

3 Stars from Doody
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Product Details

  • ISBN-13: 9780199636457
  • Publisher: Oxford University Press, USA
  • Publication date: 1/28/2002
  • Series: Practical Approach Series, #255
  • Edition description: REV
  • Edition number: 2
  • Pages: 312
  • Product dimensions: 7.30 (w) x 9.80 (h) x 0.90 (d)

Table of Contents

Preface to the first edition
Preface to the second edition
Protocols list
Abbreviations
1 Strategies for research in molecular biology 1
Strategies for gene isolation 1
Studying gene structure and expression 10
Interpretation of molecular biology experiments 12
2 Construction of genomic libraries in [lambda] and cosmid vectors 15
Overview of library construction 16
Representation 17
The choice between [lambda] and cosmid libraries 19
Vectors 19
Library construction 20
3 Construction of a cDNA library 41
Isolation of total RNA 43
Preparation of poly(A[superscript +]) RNA 44
cDNA synthesis 46
Preparation of cDNA for cloning 48
Establishment of the cDNA library 55
Characterization of the cDNA library 59
Amplification and storage of the cDNA library 61
4 Nucleic acid labelling and detection 63
Radioactive labelling and detection 69
Non-radioactive labelling and detection methods 101
5 Immobilization of nucleic acids and hybridization analysis 109
Immobilization of nucleic acids on filters 109
Hybridization analysis of immobilized nucleic acids 133
Advanced hybridization techniques 151
6 DNA sequencing 157
Preparation of template DNA 158
The sequencing reactions 166
Gel electrophoresis and autoradiography 172
Building a long contiguous sequence 179
Automated DNA sequencing 183
DNA sequence storage and analysis 184
7 The polymerase chain reaction 187
Methodology for PCR 187
Cloning PCR products 201
Sequencing PCR products 203
Problems with PCR 207
8 Protein expression in Escherichia coli 209
Factors affecting protein expression 209
Protein fusion systems 211
Expression of proteins in the periplasm 217
Cofactor requirements 219
Problems when scaling up expression 219
9 Transcript mapping 223
Mapping by nuclease protection 226
Primer-extension mapping 239
A1 Health hazards and safety procedures 245
A2 Equipment for molecular biology research 251
A3 Important Escherichia coli strains 255
A4 Recipes and general procedures 261
A5 Restriction endonucleases 267
A6 DNA and RNA modification enzymes 275
A7: List of suppliers 279
Index 285
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