RNA Methodologies

A Laboratory Guide for Isolation and Characterization


  • Robert E. Farrell, Jr., Penn State University, York, PA, U.S.A.

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.
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Researchers, graduate students and lab technicians in the fields of molecular biology, cell biology, and biochemical genetics.


Book information

  • Published: April 2005
  • ISBN: 978-0-12-249696-7


"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."
--Dr. Patrick Pagesy at INSERM for HEMOGLOBIN

Table of Contents

RNA and the Cellular Biochemistry RevisitedTranscription and the Organization of Eukaryotic GenesMessenger RNAResilient RibonucleasesRNA Isolation StrategiesThe Truth About TissuesIsolation of Polyadenylated RNAQuality Control for RNA PreparationsDot Blot AnalysisElectrophoresis of RNAPhotodocumentation and Image AnalysisNorthern AnalysisNucleic Acid Probe TechnologyPractical Nucleic Acid HybridizationPrinciples of DetectionQuantification of Specific mRNAs by Nuclease ProtectionAnalysis of Nuclear RNAcDNA SynthesisRT-PCRQuantitative PCR TechniquesTranscript Subtraction MethodsmRNA Differential DisplayHigh Throughput Analysis of Gene ExpressionRNA InterferenceGenomes, Transcriptomes, Proteomes, and BioinformaticsAn RNA Paradigm