RNA Methodologies, Third Edition: A Laboratory Guide for Isolation and Characterization

✍ Scribed by Robert E. Farrell (Author)

• Publisher: Academic Press
• Year: 2005
• Tongue: English
• Leaves: 770
• Edition: 3
• Category: Library

✦ Synopsis

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

Preface......Page 1
1. RNA and the Cellular Biochemistry Revisited......Page 4
2. Transcription and the Organization of Eukaryotic Genes......Page 20
3. Messenger RNA......Page 35
4. Resilient Ribonucleases......Page 50
5. RNA Isolation Strategies......Page 70
6. The Truth About Tissues......Page 117
7. Isolation of Polyadenylated RNA......Page 141
8. Quality Control for RNA Preparations......Page 166
9. Dot Blot Analysis......Page 182
10. Electrophoresis of RNA......Page 193
11. Photodocumentation and Image Analysis......Page 241
12. Northern Analysis......Page 264
13. Nucleic Acid Probe Technology......Page 288
14. Practical Nucleic Acid Hybridization......Page 320
15. Principles of Detection......Page 336
16. Quantification of Specific mRNAs by Nuclease Protection......Page 364
17. Analysis of Nuclear RNA......Page 390
18. cDNA Synthesis......Page 424
19. RT-PCR......Page 442
20. Quantitative PCR Techniques......Page 493
21. Transcript Subtraction Methods......Page 539
22. mRNA Differential Display......Page 558
23. High-Throughput Analysis of Gene Expression......Page 591
24. RNA Interference: Targeted Gene Silencing......Page 604
25. Genomes, Transcriptomes, Proteomes, and Bioinformatics......Page 623
26. An RNA Paradigm......Page 635
Appendix M: Electrophoresis: Principles, Parameters, and Safety......Page 647
Epilogue: A Few Pearls of Wisdom......Page 661
Appendix A: Maintaining Complete and Accurate Records......Page 668
Appendix B: Useful Stock Solutionsfor the Molecular Biologist......Page 671
Appendix C: Phenol Preparation......Page 678
Appendix D: Disposal of Ethidium Bromide and SYBR Green Solutions......Page 683
Appendix E: DNase I Removal of DNA from an RNA Sample......Page 687
Appendix F: RNase Incubation to Remove RNA from a DNA Sample......Page 689
Appendix G: Deionization of Formamide,Formaldehyde, and Glyoxal......Page 691
Appendix H: Silanizing Centrifuge Tubes and Glassware......Page 693
Appendix I: Centrifugation as a Mainstream Tool for the Molecular Biologist......Page 694
Appendix J: Trypsinization Protocol for Anchorage-Dependent Cells......Page 701
Appendix K: Isolation of High-Molecular-Weight DNA by Salting-Out......Page 704
Appendix L: RNA Isolation from Plant Tissue......Page 708
Appendix N: Polyacrylamide Gel Electrophoresis......Page 710
Appendix O: Selected Suppliers of Equipment, Reagents, and Services......Page 713
Appendix P: Useful SI Units......Page 718
Appendix Q: Common Abbreviations......Page 719
Appendix R: Trademark Citations......Page 721
Glossary......Page 725
Index......Page 742