| Description |
1 online resource (333 pages) |
| Contents |
""Book Cover""; ""Half-Title""; ""Title""; ""Copyright""; ""PCR Technology: An Introduction to Current Trends and Innovations""; ""The Editors""; ""Contributors""; ""Contents""; ""Section I General Considerations""; ""Template Isolation and Preparation""; ""1 Extraction and Amplification of Ancient DNA ""; ""I. INTRODUCTION""; ""II. MATERIALS AND METHODS""; ""A. SAMPLE PREPARATION""; ""B. PHENOL CHLOROFORM EXTRACTION PROTOCOL""; ""C. SILICA AND GUANIDINIUM THIOCYANATE EXTRACTION PROTOCOL""; ""D. PCR AMPLIFICATION""; ""III. DISCUSSION""; ""REFERENCES""; ""2 Long PCR from Damaged DNA "" |
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""I. INTRODUCTION""""II. USE OF LONG PCR TO DETECT MTDNA DAMAGE""; ""A. DNA ISOLATION, MATERIALS, AND PCR INSTRUMENTATION""; ""B. PCR PROTOCOL""; ""1. Principle of the Four-Primer Long PCR Protocol""; ""2. PCR Conditions""; ""3. Analysis of PCR Products and Interpretation of Data""; ""C. POSSIBLE PITFALLS, TROUBLESHOOTING TIPS, AND OTHER REMARKS""; ""III. USE OF EXONUCLEASE III TO ENHANCE LONG PCR AMPLIFICATION OF DAMAGED DNA TEMPLATES""; ""A. EFFECTS OF EXONUCLEASE III ON THE AMPLIFICATION OF DAMAGED MTDNA TEMPLATES"" |
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""B. EFFECTS OF EXONUCLEASE III ON THE AMPLIFICATION OF DAMAGED NUCLEAR DNA TEMPLATES""""1. Extraction of DNA, Materials, and PCR Instrumentation""; ""2. PCR Conditions""; ""3. Analysis of PCR Products and Interpretation of Data""; ""C. COMMON PITFALLS, TROUBLESHOOTING TIPS, AND SPECIFIC REMARKS""; ""IV. CONCLUSIONS""; ""REFERENCES""; ""3 Quantitative mRNA Analysis in Small Cell Samples by RT-PCR and Flow Cytometry ""; ""I. INTRODUCTION""; ""II. MATERIALS AND METHODS""; ""A. ISOLATION AND CULTURE OF ENDOTHELIAL CELLS""; ""B. ENDOTHELIAL CELL SORTING BY FLOW CYTOMETRY"" |
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""C. PREPARATION OF CELL LYSATES FOR RT-PCR""""D. COUNTING OF CELL NUCLEI BY FLOW CYTOMETRY: CALIBRATION OF THE FLOW CYTOMETER""; ""E. RT-PCR, PRIMERS, AND INTERNAL STANDARD""; ""III. RESULTS AND DISCUSSION""; ""ACKNOWLEDGMENTS""; ""REFERENCES""; ""4. Microdissection of Single or Small Numbers of Cells for Analyses of DNA and RNA by PCR ""; ""I. INTRODUCTION""; ""II. EXAMPLES""; ""A. INSTRUMENTATION AND SUPPLIERS""; ""1. Laser-Based Microdissection Systems""; ""2. Hydraulic Micromanipulation (all instruments from Narishige, Tokyo, Japan)""; ""3. Cryostat Microtome"" |
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""B. REAGENTS AND SUPPLIERS""""C. EXPERIMENTAL PROTOCOL""; ""1. Tissue Collection and Sectioning""; ""2. Staining for DNA Extraction""; ""3. Staining for RNA Extraction""; ""4. Dehydration""; ""5. Hydraulic Micromanipulation""; ""6. Laser Capture Microdissection""; ""7. Leica AS LMD Laser Microdissection""; ""8. Single-Cell PCR and DNA Sequencing""; ""D. INTERPRETATION OF DATA AND RESULTS""; ""E. COMMON PITFALLS AND TROUBLESHOOTING TIPS""; ""III. DISCUSSION""; ""A. BUDGET, EXPERTISE, AND EFFORT REQUIRED""; ""B. OTHER USEFUL APPLICATIONS OF THE METHOD""; ""C. POTENTIAL FUTURE DEVELOPMENTS"" |
| Notes |
""Acknowledgments"" |
| Bibliography |
Includes bibliographical references and index |
| Notes |
Print version record |
| Subject |
Polymerase chain reaction -- Methodology
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Polymerase chain reaction -- Methodology
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| Form |
Electronic book
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| Author |
Griffin, Hugh G
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Griffin, Annette M
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| ISBN |
9781420040654 |
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1420040650 |
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9780203010549 |
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020301054X |
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