RNA Methodologies

3rd Edition

A Laboratory Guide for Isolation and Characterization

Authors: Robert E. Farrell, Jr.
Hardcover ISBN: 9780122496967
eBook ISBN: 9780080454764
Imprint: Academic Press
Published Date: 1st April 2005
Page Count: 688
89.95 + applicable tax
72.00 + applicable tax
118.00 + applicable tax
Unavailable
Compatible Not compatible
VitalSource PC, Mac, iPhone & iPad Amazon Kindle eReader
ePub & PDF Apple & PC desktop. Mobile devices (Apple & Android) Amazon Kindle eReader
Mobi Amazon Kindle eReader Anything else

Institutional Access


Description

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.

Key Features

  • 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

Readership

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

Table of Contents

Dedication

Preface

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

Gene Organization

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

Rationale

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

Rationale

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

Other Methods

Short- and Long-Term Storage of Purified RNA

Chapter 6: The Truth About Tissues

Rationale

Tissue Culture or Tissue?

Homogenization Methods

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

Rationale

Polyadenylation

The Poly(A) Caveat

Selection of Polyadenylated Molecules

Magne

Details

No. of pages:
688
Language:
English
Copyright:
© Academic Press 2005
Published:
Imprint:
Academic Press
eBook ISBN:
9780080454764
Hardcover ISBN:
9780122496967

About the Author

Robert E. Farrell, Jr.

Affiliations and Expertise

Penn State University, York, PA, U.S.A.

Reviews

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