This book gives a comprehensive account of the practical aspects of PCR and strong consideration is given to ensure its optimal use in a laboratory environment. This includes the theoretical aspects related to PCR, setting-up of a PCR laboratory and good Laboratory Practice and standardised PCR prools to detect animal disease pathogens. Examples of Standard Operating Procedures as used in individual specialist laboratories and an outline of training materials necessary for PCR technology transfer are presented. The difficulties, advantages and disadvantages in PCR applications are explained and placed in context with other test systems. Emphasis is placed on the use of PCR for detection of pathogens with a particular focus on diagnosticians and scientists from the developing world. It is hoped that this book will enable readers from various disciplines and levels of expertise to better judge the merits of PCR and to increase their skills and knowledge in order to assist in a more logical, efficient and assured use of this technology.
|Edition description:||Softcover reprint of hardcover 1st ed. 2005|
|Product dimensions:||6.30(w) x 9.45(h) x 0.03(d)|
Table of Contents
Chapter One: Background.
Chapter Two: PCR – the basic reaction.
Chapter Three: Amplicon detection, analysis and PCR evaluation.
Chapter Four: Limitations and Troubleshooting.
Chapter Five: Specific types of PCR assays.
Chapter Six: The PCR laboratory.
Chapter Seven: Disease diagnosis using PCR specific procedures for important veterinary pathogens.
Appendices. Technical. Information and general prools.
Appendix A: Thermal cyclers.
Appendix B: Basic prool for agarose gel electrophoresis.
Appendix C: Using nucleic acid hybridisation in order to verify the integrity of a PCR amplicon.
Appendix D: Digestion of DNA with restriction endonucleases and electrophoresis of the restriction fragments.
Appendix E: DNA sequence analysis.
Appendix F: An example of a RT-PCR prool.
Appendix G (i): Calibration and Maintenance of Mechanical Pipettes.
Appendix G (ii): Standard Operating Procedure for using micropipettes.
Appendix H: Standard Operating Procedure for using clocks and timers.
Appendix I: Theoretical background to pH, buffers and concentration of solutions.
Appendix J: General buffer and sk solutions.
Appendix K: General reagents and suppliers.
Appendix L: Bioinformatics.
Original Papers. Glossary. Index.