Keywords:
Polymerase chain reaction.
;
Gene amplification.
;
Electronic books.
Description / Table of Contents:
The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual. Key Features * This first-rate guide will help you * Avoid contamination--with specific instructions on setting up your lab * Avoid cumbersome molecular biological techniques * Discover new applications * Simply call our toll-free, 800 number listed below or cut out the coupon, fill in the blanks, and mail it to us with your check, credit-card number, or money order; We'll send you the book and you'll get your answers.
Type of Medium:
Online Resource
Pages:
1 online resource (501 pages)
Edition:
1st ed.
ISBN:
9780080886718
URL:
https://ebookcentral.proquest.com/lib/geomar/detail.action?docID=1131451
DDC:
572.86
Language:
English
Note:
Front Cover -- PCR Protocols: A Guide to Methods and Applications -- Copyright Page -- Table of Contents -- Contributors -- Preface -- Part One: BASIC METHODOLOGY -- Chapter 1. Optimization of PCRs -- Standard PCR Amplification Protocol -- Enzyme Concentration -- Deoxynucleotide Triphosphates -- Magnesium Concentration -- Other Reaction Components -- Primer Annealing -- Primer Extension -- Denaturation Time and Temperature -- Cycle Number -- Primers -- Plateau Effect -- Fidelity Considerations -- Chapter 2. Amplification of Genomic DNA -- Protocols -- Other Reaction Components -- Cycling Parameters -- Chapter 3. Amplification of RNA -- Protocols -- Discussion and Helpful Hints -- Chapter 4. RACE: Rapid Amplification of cDNA Ends -- RACE Protocol -- Cloning RACE Products -- Troubleshooting and Comments on Protocol Steps -- Chapter 5. Degenerate Primers for DNA Amplification -- Recommended Procedures -- Examples of Use -- Chapter 6. cDNA Cloning Using Degenerate Primers -- Principle of the MOPAC Procedure -- Further Advances in MOPAC Technology -- Further Recommendations for the MOPAC Procedure -- Protocols -- Acknowledgments -- Chapter 7. PCR with 7-Deaza-2'-Deoxyguanosine Triphosphate -- PCR Using a 3 : 1 c7dGTP : dGTP Mixture -- Results and Discussion -- Acknowledgment -- Chapter 8. Competitive PCR for Quantitation of mRNA -- Protocols -- Example -- Discussion -- Chapter 9. Quantitative PCR -- Protocols -- Discussion -- Chapter 10. Production of Single-Stranded DNA by Asymmetric PCR -- Protocol -- Discussion -- Acknowledgments -- Chapter 11. Cloning with PCR -- Primer Design -- Protocols -- Sticky-End Cloning -- Discussion -- Chapter 12. Oligonucleotide Ligation Assay -- Protocol -- Design and Labeling of Oligonucleotides -- Discussion -- Acknowledgments -- Chapter 13. Nonisotopically Labeled Probes and Primers -- Chemistry -- Protocols.
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Summary -- Acknowledgments -- Chapter 14. Incorporation of Biotinylated dUTP -- Protocol -- Discussion -- Chapter 15. Nonisotopic Detection of PCR Products -- Protocols -- Preparation of Membranes -- Color Development -- Example of Use -- Troubleshooting -- Conclusion -- Chapter 16. Thermostable DNA Polymerases -- Characterized Thermophilic Organisms -- Thermus aquaticus -- Fidelity, Extension of Mismatched Primers, and Mutagenesis -- Template Requirements and Novel Properties -- Ionic Requirements, Solvents, and Inhibitors -- Endogenous DNA -- Acknowledgments -- Chapter 17. Procedures to Minimize PCR-Product Carry-Over -- Physical Separation of Pre- and Post-PCR Amplifications -- Aliquot Reagents -- Positive Displacement Pipettes -- Meticulous Laboratory Techniques -- Judicious Selection of Controls -- Chapter 18. Sample Preparation from Blood, Cells, and Other Fluids -- Protocols -- Discussion and Summary -- Acknowledgments -- Chapter 19. Sample Preparation from Paraffin-Embedded Tissues -- Protocols -- Discussion -- Acknowledgments -- Chapter 20. Amplifying Ancient DNA -- Protocols -- Discussion -- Acknowledgments -- Part Two: RESEARCH APPLICATIONS -- Chapter 21. In Vitro Transcription of PCR Templates -- Protocols -- Applications -- Chapter 22. Recombinant PCR -- Protocols -- Misincorporation by Taq DNA Polymerase -- Chapter 23. DNase I Footprinting -- Protocols -- Discussion -- Chapter 24. Sequencing with Taq DNA Polymerase -- Basic Sequencing Protocol -- Sequencing of PCR Products -- Acknowledgments -- Chapter 25. Direct Sequencing with the Aid of Phage Promoters -- Protocols for RAWTS -- Nomenclature -- Discussion -- Acknowledgments -- Chapter 26. Identifying DNA Polymorphisms by Denaturing Gradient Gel Electrophoresis -- Strategies for Identifying Polymorphisms by DGGE -- Protocols -- Conclusions -- Acknowledgments.
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Chapter 27. Amplification of Flanking Sequences by Inverse PCR -- Protocol for Inverse PCR -- Using Inverse PCR for Chromosome Walking -- Further Applications of Inverse PCR -- Chapter 28. Detection of Homologous Recombinants -- Basic Strategy for Screening for Homologous Recombinants -- Potential Artifacts -- Protocols -- Chapter 29. RNA Processing: Apo-B -- Protocols -- Discussion -- Chapter 30. A Transcription-Based Amplification System -- Protocols -- Results and Conclusions -- Chapter 31. Screening of λgt11 Libraries -- Protocol -- Variations -- Part Three: GENETICS AND EVOLUTION -- Chapter 32. HLA DNA Typing -- PCR/Oligonucleotide Probe Typing -- Protocols -- Potential Problems -- Chapter 33. Multiplex PCR for the Diagnosis of Duchenne Muscular Dystrophy -- Multiplex Amplification Protocol -- Discussion -- Potential Problems and Solutions -- Future Developments -- Acknowledgments -- Chapter 34. Isolation of DNA from Fungal Mycelia and Single Spores -- DNA Isolation Protocol -- Notes -- Amplification from Single Spores -- Chapter 35. Genetic Prediction of Hemophilia A -- Protocols and Examples -- Analysis of Polymorphisms by Hybridization of Allele-Specific Oligonucleotides -- Discussion -- Acknowledgments -- Chapter 36. Haplotype Analysis from Single Sperm or Diploid Cells -- Method -- Protocols -- Chapter 37. Amplification of Ribosomal RNA Genes for Molecular Evolution Studies -- Protocols -- Results and Discussion -- Conclusions -- Acknowledgment -- Chapter 38. Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics -- Protocol -- Part Four: DIAGNOSTICS AND FORENSICS -- Chapter 39. Detection of Human T-Cell Lymphoma/Leukemia Viruses -- Protocols -- Acknowledgments -- Chapter 40. Detection of Human Immunodeficiency Virus -- Protocols -- Interpretation of Results -- Applications.
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Chapter 41. Detection of Hepatitis B Virus -- Protocols -- DNA Amplification -- Chapter 42. Detection and Typing of Genital Human Papillomaviruses -- Protocols -- Analysis and Interpretation of Results -- Acknowledgments -- Chapter 43. Detection of Human Cytomegalovirus -- Protocols -- Comments -- Chapter 44. PCR Amplification of Enteroviruses -- Protocols -- Example -- Chapter 45. Novel Viruses -- Amplification of Novel Viral Sequences Using Degenerate Oligonucleotide Primers -- Chapter 46. Analysis of ras Gene Point Mutations by PCR and Oligonucleotide Hybridization -- Principle of the Method -- Protocol -- Chapter 47. B-Cell Lymphoma: t(14 -- 18) Chromosome Rearrangement -- Protocols -- Standardization of the Procedure -- Contamination -- Chapter 48. Detecting Bacterial Pathogens in Environmental Water Samples by Using PCR and Gene Probes -- Protocols -- Chapter 49. PCR in the Diagnosis of Retinoblastoma -- Protocols -- cDNA Reaction -- Discussion -- Chapter 50. Determination of Familial Relationships -- Strategy for the Amplification and Direct Sequencing of the D Loop -- Protocols -- PCR Procedure -- Population Genetics of the Human D Loop -- Determination of Maternal Relationships via mtDNA D Loop Sequencing -- Acknowledgments -- Part Five: INSTRUMENTATION AND SUPPLIES -- Chapter 51. PCR in a Teacup: A Simple and Inexpensive Method for Thermocycling PCRs -- Supplies -- Acknowledgments -- Chapter 52. A Low-Cost Air-Driven Cycling Oven -- System Overview -- Details of Construction -- Sources for Specialized Materials -- Chapter 53. Modification of a Histokinette for Use as an Automated PCR Machine -- Modification of a Histokinette -- Procedure -- Notes -- Examples of Use -- Acknowledgments -- Chapter 54. Organizing a Laboratory for PCR Work -- Preventing Cross-Contamination with Amplified DNA Sequences.
,
Estimated Cost of PCR and Direct Sequencing -- Acknowledgments -- Chapter 55. Basic Equipment and Supplies -- Index.
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