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
Researchers, graduate students and lab technicians in the fields of molecular biology, cell biology, and biochemical genetics.
Chapter 1: RNA and the Cellular Biochemistry Revisited
Why Study RNA?
What Is RNA?
Assembly of Polynucleotides
Types of RNA
Stringency: Conditions That Influence Nucleic Acid Structure
Types of Double-Stranded Molecules
Chapter 2: Transcription and the Organization of Eukaryotic Genes
Transcription and the Central Dogma
RNA Polymerases and the Products of Transcription
Chapter 3: Messenger RNA
Topology of a Typical mRNA Molecule
Stability in the Cytoplasm
Levels of Regulation
Chapter 4: Resilient Ribonucleases
Elimination of Ribonuclease Activity
Types of Ribonuclease Inhibitors
Preparation of Equipment and Reagents
Other Reagents Used to Control Nuclease Activity
Protocol: Synthesis of Vanadyl Ribonucleoside Complexes
Chapter 5: RNA Isolation Strategies
Goals in the Purification of RNA
Lysis Buffer Formulations
Isolation of RNA with Guanidinium Buffers
Guanidinium-Acid-Phenol Extraction Techniques
Density Gradient Centrifugation
Simultaneous Isolation of RNA and DNA
The Word on Kits
Short- and Long-Term Storage of Purified RNA
Chapter 6: The Truth About Tissues
Tissue Culture or Tissue?
RNA Isolation Strategies for Various Organs and Tissues
Protocol: LiCl–Urea Method for RNA Isolation from Tissue
Protocol: RNA Isolation from Lipid-Enriched Tissue
Purification of Polysome-Engaged mRNA
Collecting Samples in the Field
RNA “Clean-Up” Methods
Chapter 7: Isolation of Polyadenylated RNA
The Poly(A) Caveat
Selection of Polyadenylated Molecules
- No. of pages:
- © Academic Press 2005
- 1st April 2005
- Academic Press
- eBook ISBN:
- Hardcover ISBN:
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.
Penn State University, York, PA, USA
Praise for the third edition @qu:"This third edition of RNA Methodologies is an updated version of a laboratory guide edited for the first time in 1993; it represents an exhaustive collection of tried, tested, and optimized protocols for isolation and chracterization of RNA. The third edition of RNA Methodologies is a well-documented guide for any researcher who has to manage RNA studies. Each chapter of this book provides a rationale to help in the decision-making process...The inclusion of troubleshooting hints and annotations based on personal experience are provided to improve the method(s). This laboratory guide offers protocols on traditional and new technologies, with a focus on how to use the expedient technique for achieving an experimental goal in the study of gene expression." @source:--Dr. Patrick Pagesy at INSERM for HEMOGLOBIN