RNA Methodologies

Laboratory Guide for Isolation and Characterization

By

  • Robert E. Farrell, Jr., Penn State University, York, PA, U.S.A.

This is the fourth edition of the successful laboratory guide which has translated the rich story of riboneucleic acid for over fifteen years. RNA Methodologies 4e presents the latest collection of tested laboratory protocols for the isolation and characterization of eukaryotic and prokaryotic RNA with greater emphasis on transcript profiling, including quantification issues and elucidation of alternative transcription start sites. Collectively the chapters work together providing analysis with clear take-home lessons to assist researchers to understand RNA and to optimize time at the bench. The abundant use of flow charts, tables and graphs are especially helpful in the planning and implementation phases of a project and facilitate learning. 30% new material in this edition includes the addition of RNA isolation protocols including RNA isolation from tissue, expansion of PCR optimization analysis and RNA interference sections, the introduction of a new chapter dealing with the molecular biology of plants, and an expanded glossary.
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Audience

Researchers, graduate students and lab technicians in the fields of molecular biology, cell biology, and biochemical genetics

 

Book information

  • Published: September 2009
  • Imprint: ACADEMIC PRESS
  • ISBN: 978-0-12-374727-3


Table of Contents

**Fully revised with five new chapters, a new appendix, and nine chapters with major expansions**

CHAPTER 1: RNA and the Cellular Biochemistry Revisited
CHAPTER 2: RNA Isolation Strategies
CHAPTER 3: The Truth about Tissues
CHAPTER 4: Going Green: RNA and the Molecular Biology of Plants (NEW)
CHAPTER 5: Isolation of Polyadenylated RNA
CHAPTER 6: Quality Control for RNA Preparations (EXPANDED)
CHAPTER 7: Resilient Ribonucleases
CHAPTER 8: Stringency: Conditions that Influence Nucleic Acid Structure (EXPANDED)
CHAPTER 9: Electrophoresis of RNA (EXPANDED)
CHAPTER 10: Photodocumentation and Image Analysis (EXPANDED)
CHAPTER 11: Northern Analysis
CHAPTER 12: Nucleic Acid Probe Technology
CHAPTER 13: Practical Nucleic Acid Hybridization
CHAPTER 14: Principles of Detection (EXPANDED)
CHAPTER 15: Quantification of Specific mRNAs by Nuclease Protection
CHAPTER 16: Analysis of Nuclear RNA (EXPANDED)
CHAPTER 17: cDNA Synthesis (EXPANDED)
CHAPTER 18: RT-PCR (NEW)
CHAPTER 19: Quantitative PCR Techniques
CHAPTER 20: Functional Genomics and Transcript Profiling (NEW)
CHAPTER 21: High-Throughput Analysis of Gene Expression
CHAPTER 22: Non-Array Methods for Global Analysis of Gene Expression (NEW)
CHAPTER 23: RNA Interference: Targeted Gene Silencing (EXPANDED)
CHAPTER 24: Genomes, Transcriptomes, Proteomes, and Bioinformatics
CHAPTER 25: An RNA Paradigm

APPENDIX A: Maintaining Complete and Accurate Records
APPENDIX B: Converting Mass to Moles (NEW)
APPENDIX C: Useful Stock Solutions for the Molecular Biologist
APPENDIX D: Phenol Preparation
APPENDIX E: Disposal of Ethidium Bromide and SYBR Green Solutions
APPENDIX F: DNase I Removal of DNA From An RNA Sample
APPENDIX G: DNase Incubation to Remove RNA from a DNA Sample
APPENDIX H: Deionization of Formamide, Formaldehyde, and Glyoxal
APPENDIX I: Silanizing Centrifuge Tubes and Glassware
APPENDIX J: Trypsinization Protocol for Anchorage-Dependent Cells
APPENDIX K: Isolation of High-Molecular-Weight DNA by Salting-Out
APPENDIX L: Electrophoresis: Principles, Parameters, and Safety
APPENDIX M: Polyacrylamide Gel Electrophoresis
APPENDIX N: Dot Blot Analysis (NEW)
APPENDIX O: Centrifugation as a Mainstream Tool for the Molecular Biologist
APPENDIX P: Selected Suppliers of Equipment, Reagents, and Services
APPENDIX Q: Useful SI Units
APPENDIX R: Common Abbreviations
APPENDIX S: Trade Citations