A Low-Cost Approach to PCR
Appropriate Transfer of Biomolecular Techniques
Published on 27. May 1999
Book
Paperback/Softback
328 pages
978-0-19-511926-8 (ISBN)
Description
The polymerase chain reaction (PCR) is a technique used to replicate specific pieces of DNA millions of times, which permits the detection and analysis of minute amounts of nucleic acids. Since its introduction in the late 1980s, this technique has been applied not only in molecular biology research but also in fields as diverse as anthropology, phylogeny, and forensics. However, despite the large impact of PCR, many of its applications remain within the confines of research and the academic environment. Now, in A Low-Cost Approach to PCR: Appropriate Transfer of Biomolecular Techniques, Dr. Eva Harris makes this elegantly simple technique more accessible to researchers, physicians, and laboratory workers throughout the world. She provides a description of the theoretical basis of the technique, the practical details of the method, and the philosophy behind the technology transfer program that she developed over the last ten years. The book serves as a guide for potential users in developing countries and for scientists in developed countries who may wish to work abroad. In addition, the low-cost approach outlined in this book can be useful for high school, undergraduate, or continuing education programs in the United States. While the specific applications of PCR outlined in the book are immediately useful to the study of infectious diseases, the approach presented can be generalized to a number of other technologies and situations. The book will help laboratories in many areas of the world generate information on site for use by physicians, epidemiologists, public health workers, and health policy professionals to develop new strategies for disease control.
Reviews / Votes
"Makes the polymerase chain reaction (PCR) technique more accessible to researchers, physicians, and laboratory workers throughout the world. Describes the theoretical basis of the technique, practical details, and the philosophy behind the technology transfer program that the author has developed over the past 10 years. A low-cost approach is outlined for use in the study of infectious diseases, which can be applied to other technologies and applications. Thisapproach is especially useful for laboratories in developing countries, and in high school undergraduate and continuing education programs in the US. Appendices give information on construction of
equipment, and contain sample charts and worksheets. Includes b&w photos of equipment, and of men and women performing lab procedures. The author is on the faculty of the School of Public Health at UC-Berkeley."--SciTech Book News
"The polymerase chain reaction (PCR) is a technique used to replicate specific pieces of DNA millions of times, which permits the detection and analysis of minute amounts of nucleic acids. . . . [T]his technique has been applied not only in molecular biology research but also in fields as diverse as anthropology, phylogeny and forensics. However, despite the large impact of PCR, many of its applications remain within the confines of research and the academic
environment. This book attempts to make the PCR technique more accessible to researchers, physicians and laboratory workers throughout the world by providing a description of the theoretical basis of the
technique, practical details of the method, and the philosophy behind the technology transfer programme that the author has developed over the last 10 years. . . . The book should help laboratories in many areas of the world generate information on site for use by physicians, epidemiologists, public health workers . . ."--CAB Abstracts
"Makes the polymerase chain reaction (PCR) technique more accessible to researchers, physicians, and laboratory workers throughout the world. Describes the theoretical basis of the technique, practical details, and the philosophy behind the technology transfer program that the author has developed over the past 10 years. A low-cost approach is outlined for use in the study of infectious diseases, which can be applied to other technologies and applications. This
approach is especially useful for laboratories in developing countries, and in high school undergraduate and continuing education programs in the US. Appendices give information on construction of
equipment, and contain sample charts and worksheets. Includes b&w photos of equipment, and of men and women performing lab procedures. The author is on the faculty of the School of Public Health at UC-Berkeley."--SciTech Book News
"The polymerase chain reaction (PCR) is a technique used to replicate specific pieces of DNA millions of times, which permits the detection and analysis of minute amounts of nucleic acids. . . . [T]his technique has been applied not only in molecular biology research but also in fields as diverse as anthropology, phylogeny and forensics. However, despite the large impact of PCR, many of its applications remain within the confines of research and the academic
environment. This book attempts to make the PCR technique more accessible to researchers, physicians and laboratory workers throughout the world by providing a description of the theoretical basis of the
technique, practical details of the method, and the philosophy behind the technology transfer programme that the author has developed over the last 10 years. . . . The book should help laboratories in many areas of the world generate information on site for use by physicians, epidemiologists, public health workers . . ."--CAB Abstracts
More details
Language
English
Place of publication
New York
United States
Target group
Professional and scholarly
Illustrations
halftones
Dimensions
Height: 234 mm
Width: 156 mm
Thickness: 18 mm
Weight
500 gr
ISBN-13
978-0-19-511926-8 (9780195119268)
Copyright in bibliographic data and cover images is held by Nielsen Book Services Limited or by the publishers or by their respective licensors: all rights reserved.
Schweitzer Classification
Other editions
Additional editions
E-Book
12/1998
1st Edition
OUP eBook
€108.99
Available for download
Person
Content
1 INTRODUCTION ; 2 PCR TECHNOLOGY ; 2.1 Description of the Technique ; 2.1.1 Historical Overview ; 2.1.2 The Molecular Basis of PCR ; 2.1.3 Advantages and Disadvantages ; 2.2 Technical Details ; 2.2.1 Components of the PCR Mixture ; 2.2.2 Thermal Cycling Parameters ; 2.2.3 Optimizing ; 2.2.4 Potential Problems ; 2.2.5 Detection of Products ; 2.3 Frequently-Used PCR-Based Techniques ; 3 PRINCIPLES OF SUSTAINABLE TECHNOLOGY TRANSFER ; 3.1 A Low-Cost Methodology ; 3.1.1 Appropriate Technology ; 3.1.2 Alternative Techniques ; 3.1.3 Simplification of Protocols ; 3.1.4 In-House Preparation of Reagents ; 3.1.5 Recycling ; 3.1.6 Donated Materials ; 3.2 Knowledge-Based Participatory Transfer Process ; 3.3 Appropriate Application ; 3.3.1 General Considerations ; 3.3.2 An Evaluation Framework ; 3.3.3 Assessment Criteria for PCR ; 3.3.4 Case-by-Case Evaluation of PCR Applied to Infectious Diseases ; 3.4 Intra-Reginal Cooperation ; 4 CASE STUDY: THE AMB/ATT PROGRAM ; 4.1 Introduction ; 4.2 Program Description ; 4.2.1 Objectives ; 4.2.2 Format ; 4.3 Program Structure ; 4.3.1 Phase I ; 4.3.2 Phase II ; 4.3.3 Phase III ; 4.4 Program Development ; 4.5 Examples of Projects ; 4.6 International Courses ; II SELECTED PROTOCOLS ; 5.1 PCR Protocols ; 5.1.1 Overview ; 5.1.2 Equipment, Materials, Control, and Procedures Common to All PCR Protocols ; 5.1.3 Dengue Virus ; 5.1.4 New World Leishmania ; 5.1.5 Mycobactrium tuberculosis ; 5.1.6 Plasmodium falciparum and Plasmodium vivax ; 5.1.7 Vibrio cholerae ; 5.1.8 Diarrheagenic E. coli and Shigella ; 5.1.9 Chlamydia trachomatis and Neisseria gonorrhoeae ; 5.1.10 Leptospira ; 5.1.11 Trypanosoma cruzi ; 5.2 Nonradioactive DNA Probes: V. cholerae Colony Blot ; 5.2.1 Labeling the Probe Using PCR ; 5.2.2 Preparation of the Colony Blot ; 5.2.3 Hybridization ; 5.2.4 Visualization ; 6 RAPID CLONING OF PCR PRODUCTS ; 6.1 Primer Design ; 6.2 Preparation of PCR Products for Cloning ; 6.2.1 Digestion of Vector and PCR Products ; 6.2.2 Preparative Agarose Gel ; 6.2.3 Purification of the Excised DNA Fragments Using Silica Particles ; 6.3 Ligation ; 6.3.1 Checking DNA Fragment Concentration by Agarose Gel Electrophoresis ; 6.3.2 Ligation Reaction ; 6.4 Preparation of Competent Cells and Transformation ; 6.4.1 Preparation of Competent Cells ; 6.4.2 Transformation ; 6.5 Checking Clones by PCR ; 6.6 Plasmid Purification ; 6.7 Analysis of Clones by Restriction Enzyme Digestion ; 6.8 Agarose Gel Electrophoresis ; III APPENDIX ; A. CONSTRUCTION OF LABORATORY EQUIPMENT ; B. IN-HOUSE PREPARATION OF REAGENTS ; B.1 USEFUL FORMULAS ; B.2 SOLUTIONS ; B.3 PREPARATION OF SELECTED REAGENTS ; B.4 DNA SIZE MARKERS ; C. INVENTORY FOR A PCR LABORATORY ; D. GOOD LABORATORY PRACTICE ; D.1 GENERAL TIPS ; D.2 CALIBRATION OF ADJUSTABLE PIPETTORS ; E. PREVENTION OF CROSS-CONTAMINATION ; F. PCR TROUBLESHOOTING GUIDE AND FLOW-CHART ; G. WORKSHOP ORGANIZATION AND TEACHING TIPS ; H. SAMPLE CHARTS AND WORKSHEETS ; I. USEFUL WORLD WIDE WEB SITES