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By: Elizabeth van Pelt-Verkuil, Alexander van Belkum and John P Hays
Kary Mullis was awarded a Nobel prize for inventing the PCR technique more than a decade ago in 1993. Since its "discovery", multiple adaptations and variations of the standard PCR technique have been described, with many of these adaptations and variations currently being used in clinical diagnostic and research laboratories across the world, from academic laboratories purely interested in basic research to diagnostic laboratories using high throughput routine PCR testing methodologies to detect (for example) minimum residual disease and the presence/absence of particular pathogens.
Frequently, PCR technicians limit their understanding of PCR to that particular methodology they are currently familiar with. However, this approach limits their understanding and appreciation of the range of versatile PCR techniques currently available. Techniques which may be applicable, and indeed more suitable, to their own laboratory situation. This publication aims to provide the reader with a guide to the standard PCR technique and its many available variants, with particular emphasis being placed on the role of these PCR techniques in the clinical diagnostic laboratory (the central theme of this book).
As such, many important technical issues have been included in the book, including types of PCR template material, PCR optimization, the analysis of PCR products, quality control and quality assurance, variants and adaptations of the standard PCR protocol, quantitative PCR and in situ PCR.
Chapter 1. The Polymerase Chain Reaction Chapter 2. A Brief Comparison Between In Vivo DNA Replication And In Vitro PCR Amplification Chapter 3. The PCR In Practice Chapter 4. The Different Types And Varieties Of Nucleic Acid Target Molecules Chapter 5: PCR Primers Chapter 6. Deoxynucleotide Triphosphates And Buffer Components Chapter 7. Taq And Other Thermostable DNA Polymerases Chapter 8. Important Considerations For Typical, Quantitative And Real-Time PCR Protocols Chapter 9. Analysis Of PCR Amplification Products. Chapter 10. Ensuring PCR Quality - Laboratory Organisation, Pcr Optimisation And Controls. Chapter 11. Ensuring PCR Quality-Quality Criteria And Quality Assurance. Chapter 12. Variants And Adaptations Of The Standard Pcr Protocol Chapter 13. In Situ PCR Amplification (Isa) - Major Considerations, Sample Processing And Applications
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