One: Methodology.- I. Basic Methodology.- 1. Manipulation of DNA by PCR.- 2. Cloning PCR Products.- 3. Optimization of Multiplex PCRs.- 4. Preparation of Nucleic Acids for Archival Material.- 5. PCR Amplification of Viral DNA and Viral Host Cell mRNAs in Situ.- II. Quantitation.- 6. Quantitative PCR: An Overview.- 7. Quantification of DNAs by the Polymerase Chain Reaction Using an Internal Control.- 8. RT-PCR and mRNA Quantitation.- 9. Analysis of Human T-Cell Repertoires by PCR.- III. Nonisotopic Detection.- 10. Ultrasensitive Nonradioactive Detection of PCR Reactions: An Overview.- 11. Fluorescent Detection Methods for PCR Analysis.- 12. Enzyme-Labeled Oligonucleotides.- 13. Application of the Hybridization Protection Assay (HPA) to PCR.- IV. Instrumentation.- 14. PCR Instrumentation: Where Do We Stand?.- 15. Rapid Cycle DNA Amplification.- 16. Automating the PCR Process.- V. Sequencing.- 17. PCR and DNA Sequencing.- 18. Phage Promoter-Based Methods for Sequencing and Screening for Mutations.- 19. Capture PCR: An Efficient Method for Walking Along Chromosomal DNA and cDNA.- Two: Applications.- I. General Applications.- 20. In Vitro Evolution of Functional Nucleic Acids: High-Affinity RNA Ligands of the HIV-1 rev Protein.- 21. The Application of PCR to Forensic Science.- 22. Recreating the Past by PCR.- 23. Nonbiological Applications.- II. Genetic Analysis.- 24. RT-PCR and Gene Expression.- 25. Fingerprinting Using Arbitrarily Primed PCR: Application to Genetic Mapping, Population Biology, Epidemiology, and Detection of Differentially Expressed RNAs.- 26. Genetics, Plants, and the Polymerase Chain Reaction.- III. Assessment of Therapy Effectiveness.- 27. PCR Assessment of the Efficacy of Therapy in Philadelphia Chromosome-Positive Leukemias.- 28. The Detection of Minimal Residual Disease (MRD) in Acute Lymphoblastic Leukemia Using Clone-Specific Probes Directed against V(D)J Junctional Sequences.- 29. Assessment of Therapy Effectiveness: Infectious Disease.- 30. Gene Therapy.- IV. Diagnostics.- 31. PCR and Cancer Diagnostics: Detection and Characterization of Single Point Mutations in Oncogenes and Antioncogenes.- 32. Clinical Applications of the Polymerase Chain Reaction.- 33. Infectious Diseases.- Three: PCR and the World of Business.- 34. PCR in the Marketplace.- 35. PCR and Scientific Invention: The Trial of DuPont vs. Cetus.
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Edited by the inventor of PCR and two prominent experts in PCR techniques, this first comprehensive handbook on the Polymerase Chain Reaction has the most up to date methodological protocols from the world's leading laboratories. Included are exciting new techniques and enhanced methods, previously unavailable in book form, that show the novice and experienced PCR user exactly how they can optimize their results. The applications chapters are quite unique, with the foremost researchers providing not only protocols, but explaining why PCR has revolutionized their particular field. Future enhancements of PCR as well as new potential uses are discussed. Readers will learn how PCR has changed the face of diagnostic testing, cancer research, genetics, forensics, plant biology, DNA sequencing, gene therapy, and much more! Nearly forty chapters have been extensively reviewed and checked for accuracy and breadth of subject matter.
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