RNA Methodologies

A Laboratory Guide for Isolation and Characterization

Robert E. Farrell Jr., Robert E. Farrell, Jr.

• Publisher: Academic Press
• Year: 2010
• eBook ISBN: 9780080454764
• Edition: 3
• Topic: Biological Sciences
• Subtopic: Genetics & Genomics
• Index: Biological Sciences

This laboratory guide represents a growing collection of tried, tested and optimized laboratory protocols for the isolation and characterization of eukaryotic RNA, with lesser emphasis on the characterization of prokaryotic transcripts. Collectively the chapters work together to embellish the RNA story, each presenting clear take-home lessons, liberally incorporating flow charts, tables and graphs to facilitate learning and assist in the planning and implementation phases of a project.RNA Methodologies, 3rd edition includes approximately 30% new material, including chapters on the more recent technologies of RNA interference including: RNAi; Microarrays; Bioinformatics. It also includes new sections on: new and improved RT-PCR techniques; innovative 5' and 3' RACE techniques; subtractive PCR methods; methods for improving cDNA synthesis.* Author is a well-recognized expert in the field of RNA experimentation and founded Exon-Intron, a well-known biotechnology educational workshop center * Includes classic and contemporary techniques * Incorporates flow charts, tables, and graphs to facilitate learning and assist in the planning phases of projects

Table of contents

• Cover
• Title page
• Table of Contents
• Copyright
• Dedication
• Preface
• Chapter 1: RNA and the Cellular Biochemistry Revisited
• Chapter 2: Transcription and the Organization of Eukaryotic Genes
• Chapter 3: Messenger RNA
• Chapter 4: Resilient Ribonucleases
• Chapter 5: RNA Isolation Strategies
• Chapter 6: The Truth About Tissues
• Chapter 7: Isolation of Polyadenylated RNA
• Chapter 8: Quality Control for RNA Preparations
• Chapter 9: Dot Blot Analysis
• Chapter 10: Electrophoresis of RNA
• Chapter 11: Photodocumentation and Image Analysis
• Chapter 12: Northern Analysis
• Chapter 13: Nucleic Acid Probe Technology
• Chapter 14: Practical Nucleic Acid Hybridization
• Chapter 15: Principles of Detection
• Chapter 16: Quantification of Specific mRNAs by Nuclease Protection
• Chapter 17: Analysis of Nuclear RNA
• Chapter 18: cDNA Synthesis
• Chapter 19: RT-PCR
• Chapter 20: Quantitative PCR Techniques
• Chapter 21: Transcript Subtraction Methods
• Chapter 22: mRNA Differential Display
• Chapter 23: High-Throughput Analysis of Gene Expression
• Chapter 24: RNA Interference: Targeted Gene Silencing
• Chapter 25: Genomes, Transcriptomes, Proteomes, and Bioinformatics
• Chapter 26: An RNA Paradigm
• Epilogue: A Few Pearls of Wisdom
• Appendix A: Maintaining Complete and Accurate Records
• Appendix B: Useful Stock Solutions for the Molecular Biologist
• Appendix C: Phenol Preparation
• Appendix D: Disposal of Ethidium Bromide and SYBR Green Solutions
• Appendix E: DNase I Removal of DNA from an RNA Sample
• Appendix F: RNase Incubation to Remove RNA from a DNA Sample
• Appendix G: Deionization of Formamide, Formaldehyde, and Glyoxal
• Appendix H: Silanizing Centrifuge Tubes and Glassware
• Appendix I: Centrifugation as a Mainstream Tool for the Molecular Biologist
• Appendix J: Trypsinization Protocol for Anchorage-Dependent Cells
• Appendix K: Isolation of High-Molecular-Weight DNA by Salting-Out
• Appendix L: RNA Isolation from Plant Tissue
• Appendix M: Electrophoresis: Principles, Parameters, and Safety
• Appendix N: Polyacrylamide Gel Electrophoresis
• Appendix O: Selected Suppliers of Equipment, Reagents, and Services
• Appendix P: Useful SI Units
• Appendix Q: Common Abbreviations
• Appendix R: Trademark Citations
• Glossary
• Index