RNA Methodologies, Fifth Edition continues its tradition of excellence in providing the most up-to-date ribonucleic acid lab techniques for seasoned scientists and graduate students alike. This edition features new material on the exploding field of microRNA as well as the methods for the profiling of gene expression, both which have changed considerably in recent years. As a leader in the field, Dr. Farrell provides a wealth of knowledge on the topic of RNA while also giving readers helpful hints from his own personal experience in this subject area. Beginning with the most contemporary, RNA Methodologies, Fifth Edition, presents the essential techniques to use when working with RNA for the experienced practitioner while at the same time providing images and examples to aid the beginner in fully understanding this important branch of molecular biology. The next generation of scientists can look to this work as a guide for ensuring high productivity and highly representative data, as well as best practices in troubleshooting laboratory problems when they arise.
- Features new material in miRNA, MIQE guidelines, biomarkers, RNA sequencing, digital PCR and more
- Includes expanded coverage on quantitative PCR techniques, RNAi, bioinformatics, the role of locked nucleic acids, aptamer biology, PCR arrays, and other modern technologies
- Presents comprehensive, cutting-edge information covering all aspects of working with RNA
- Builds from basic information on RNA techniques to in-depth protocols to guidance on how to modify and adjust each step of a particular application
- Presents multiple avenues for addressing the same experimental goals
1. RNA and the Cellular Biochemistry Revisited 2. Creating a Ribonuclease-free Environment 3. RNA Isolation Strategies 4. The Truth about Tissues 5. Going Green: RNA and the Molecular Biology of Plants 6. Quality Control for RNA Preparations 7. cDNA: A Permanent Biochemical Record of the Cell 8. RT-PCR: A Science and an Art Form 9. Quantitative PCR Techniques 10. miRNA 11. RNA Interference
Take a RISC: Role the Dicer 12. Stringency: Conditions that Influence Nucleic Acid Structure 13. Electrophoresis of RNA 14. Photodocumentation and Image Analysis 15. Northern Analysis 16. Nucleic Acid Probe Technology 17. Isolation of Polyadenylated RNA 18. Quantification of Specific mRNAs by Nuclease Protection 19. Analysis of Nuclear RNA 20. Non-Array Methods for Global Analysis of Gene Expression 21. Genomes, Transcriptomes, Proteomes, and Bioinformatics 22. RNA Biomarker Development 23. High-Throughput Analysis of Gene Expression 24: Functional Genomics and Transcript Profiling 25. A Few RNA Success Stories
Appendix A. Maintaining Complete and Accurate Records B. Converting Mass to Moles C. Disposal of Ethidium Bromide and SYBR Green Solutions D. DNase I Removal of DNA from an RNA Sample E. RNase Incubation to Remove RNA from a DNA Sample F. Useful Stock Solutions for the Molecular Biologist G. Silanizing Centrifuge Tubes and Glassware H. Dot Blot Analysis I. Electrophoresis: Principles, Parameters, and Safety J. Polyacrylamide Gel Electrophoresis K. Centrifugation as a Mainstream Tool for the Molecular Biologist L. Trypsinization Protocol for Anchorage-Dependent Cells M. Isolation of High-Molecular-Weight DNA by Salting-Out N. Phenol Preparation O. Deionization of Formamide, Formaldehyde, and Glyoxal P. Selected Suppliers of Equipment, Reagents, and Services Q. Useful SI Units R. Common Abbreviations S. Trademark Citations
Dr. Robert Farrell is a bench-current scientist who has 35 years of experience working with RNA in the study of transcriptional and posttranscriptional regulation of gene expression in a variety of model systems. He is also experienced in animal cell culture methods. Prior to joining the faculty at Penn State University, he operated a biotech education and service firm, winning the 1998 Small Business Contractor of the Year award from the U.S. Department of Agriculture. He is the recipient of campus- and college-wide awards for excellence in teaching, and has extensive experience running RNA and specialized biotechnology hands-on laboratory training programs all over the world. He often serves as a consultant within the pharmaceutical and biotech industries. Dr. Farrell received his Ph.D. and M.S. degrees from The Catholic University of America and his B.S. in Biology from Providence College. Dr. Farrell currently serves as the campus academic officer at Penn State York.