RNA Methodologies: A Laboratory Guide for Isolation and Characterization, Sixth Edition provides the most up-to-date ribonucleic acid lab techniques for seasoned scientists and graduate students alike. This edition features new material on RNA sequencing, RNA in Situ Hybridization, non-coding RNAs, computational RNA biology, transcriptomes and bioinformatics, along with the latest advances in methods and protocols across the field of RNA investigation. As a leader in the field, Dr. Farrell provides a wealth of knowledge on the topic of RNA biology while also giving readers helpful hints and troubleshooting techniques from his own personal experience in this subject area.
This book presents the essential knowledge and techniques to use when working with RNA for the experienced practitioner, while also aiding the beginner in fully understanding this important branch of molecular biology.
1. RNA and the cellular biochemistry revisited 2. Creating a ribonuclease-free environment 000 3. Stringency: conditions that influence nucleic acid structure 4. Ribonucleic acid isolation strategies 5. Isolation of polyadenylated RNA 6. The truth about tissues 7. Going green: RNA and the molecular biology of plants 8. Quality control for RNA preparations 9. cDNA: a permanent biochemical record of the cell 10. RT-PCR: a science and an art form 11. Quantitative polymerase chain reaction techniques 12. miRNA and other noncoding RNAs 13. RNA interference and gene editing 14. Electrophoresis of RNA 15. Photodocumentation and image analysis 16. Northern analysis 17. Nucleic acid probe technology 18. Quantification of specific mRNAs by nuclease protection 19. Analysis of nuclear RNA 20. RNA in situ hybridization 21. Array analysis of gene expression 22. Subtractive and nonsubtractive methods for the analysis of gene expression 23. Transcriptomes and bioinformatics 24. RNA-seq: the premier transcriptomics tool 25. RNA biomarker discovery and validation 26. Functional genomics strategies 27. A few RNA success stories Epilogue Appendix A: Maintaining complete and accurate records Appendix B: Converting mass to moles Appendix C: Removal of DNA from an RNA sample Appendix D: Removal of RNA from a DNA sample Appendix E: Electrophoresis: principles, parameters, and safety Appendix F: Disposal of ethidium bromide and SYBR Green solutions 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: Dot blot analysis Appendix K: Useful stock solutions for the molecular biologist Appendix L: Genomes and proteomes Appendix M: Common SI prefixes Appendix N: Common abbreviations Appendix O: Select suppliers of equipment, reagents, and services Appendix P: Trademark citations Glossary
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.
