A Low-Cost Approach to PCR: Appropriate Transfer of Biomolecular TechniquesThe 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. |
Contents
II SELECTED PROTOCOLS | 87 |
Construction of Laboratory Equipment | 223 |
InHouse Preparation of Reagents | 243 |
Inventory for a PCR Laboratory | 256 |
Good Laboratory Practice | 261 |
Prevention of CrossContamination | 264 |
PCR Troubleshooting Guide and Flowchart | 267 |
Workshop Organization and Teaching Tips | 274 |
Sample Charts and Worksheets | 278 |
Useful World Wide Web Sites | 285 |
Glossary | 290 |
Afterword | 297 |
Index | 299 |
Other editions - View all
A Low-Cost Approach to PCR: Appropriate Transfer of Biomolecular Techniques Eva Harris Limited preview - 1998 |
Common terms and phrases
10X Buffer 20 mM total agarose gel AMB/ATT analysis application assay cells Centrifuge the tubes Chlamydia trachomatis Clin coli color-coded pipettor containing cycle ddH2O denaturation dengue virus detection diagnosis dNTPs epidemiology equipment ethidium bromide fragment gel electrophoresis gene genetic genome Grey Area hybridization ice bucket identification Incubate infections ingredients labeled tube laboratory Leishmania Leptospira membrane method MgCl2 Microbiol microcentrifuge tubes mineral oil minutes molecular biology mosquitoes Mycobacterium tuberculosis negative control nucleic acid nucleotides parasites pathogenic PCR amplification PCR Master Mix PCR products PCR reaction PCR-based Phase pipet tips Plasmodium falciparum plastic polymerase chain reaction polymorphism preparing the PCR primers probe procedure protocol reagents restriction enzyme reverse transcriptase RT-PCR sample sensitivity sequence silica particles solution specific strains supernatant Taq polymerase techniques temperature template thermocycler total dNTPs Transfer Trypanosoma cruzi vector Vibrio cholerae volume vortex water bath White Area Worksheet workshop μΜ
Popular passages
Page 30 - Frohman, MA, Dush, MK, and Martin, GR (1988) Rapid production of fulllength cDNAs from rare transcripts: Amplification using a single gene-specific oligonucleotide primer. Proc. Natl. Acad. Sci. USA 85, 8998-9002.
Page 30 - Livak, KJ, Flood, SJ, Marmaro, J., Giusti, W., and Deetz, K. (1995) Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization.
Page 30 - Haase, AT, Retzel, EF, and Staskus, KA (1990) Amplification and detection of lentiviral DNA inside cells. Proc. Natl. Acad. Sci. USA 87, 4971-4975.
Page 194 - HP 1994. Plasmodium falciparum: extensive polymorphism in merozoite surface antigen 2 alleles in an area with endemic malaria in Papua New Guinea. Exp. Parasitol. 79: 106-116.
Page 196 - O. (1992). Use of polymerase chain reaction for detection of toxigenic Vibrio cholerae ol strains from the Latin American cholera epidemic.
Page 195 - Identification of the four human malaria species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections. Mol. Biochem. Parasitol. 58, 283-292.
Page 58 - Brown. KN (1993) Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections.
Page 193 - Van Embden, JD, Cave, MD, Crawford, JT, Dale, JW, Eisenach, KD, Gicquel, B., Hermans, P., Martin, C., McAdam, R., Shinnick, TM, and Small, PM (1993) Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: Recommendations for a standardized methodology.
Page 222 - Vogelstein, B. and Gillespie, D. (1979) Preparative and analytical purification of DNA from agarose.
References to this book
Populations and Genetics: Legal and Socio-Ethical Perspectives Bartha Maria Knoppers Limited preview - 2003 |