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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

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