Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S Gerstein opyright◎2001 ISBNS:0-471-37972-7( Paper);0-471 (Electronic) 8 RNA Purification Lori A. Martin, Tiffany. Smith, Dawn Obermoeller Brian Bruner Martin Kracklauer and Subramanian Dharmaraj Selecting a Purification Strategy Do Your Experiments Require Total RNa or mRNA? 198 Is It possible to predict the total rna yield from a Certain mass of tissue or number of cells? 20 Is There Protein in Your RNA Preparation, and If So, Should You be concerned 202 Is Your RNa Physically Intact? Does It Matter? Which Total RNA Isolation Technique Is Most Appropriate for Your research? What Protocol modifications should Be used for RNA Isolation from difficult tissues? Is a One-Step or Two-Step mRNA(poly (A)RNA)- Purification Strategy Most Appropriate for Your Situation? 209 How Many Rounds of Oligo(dT)-Cellulose Purification Are required? 210 Which Oligo(dT)-Cellulose Format Is Most 210 Can Oligo(dT)-Cellulose Be Regenerated and Reused?.. 211 Can a Kit Designed to Isolate mRNA Directly from he Biological Sample Purify mRNA from Total RNA?... 212 Maximizing the Yield and Quality of an RNA Preparation.. 212 What Constitutes“ RNase- Free Technique”?197 8 RNA Purification Lori A. Martin, Tiffany J. Smith, Dawn Obermoeller, Brian Bruner, Martin Kracklauer, and Subramanian Dharmaraj Selecting a Purification Strategy . . . . . . . . . . . . . . . .. . . . . . . . . 198 Do Your Experiments Require Total RNA or mRNA? . . . . . 198 Is It Possible to Predict the Total RNA Yield from a Certain Mass of Tissue or Number of Cells? . . . . . . . . 201 Is There Protein in Your RNA Preparation, and If So, Should You Be Concerned? . . . . . . . . . . . . . . . . . . . . 202 Is Your RNA Physically Intact? Does It Matter? . . . . . . . . . . 202 Which Total RNA Isolation Technique Is Most Appropriate for Your Research? . . . . . . . . . . . . . . . . . . . . . 203 What Protocol Modifications Should Be Used for RNA Isolation from Difficult Tissues? . . . . . . . . . . . . . . . . 207 Is a One-Step or Two-Step mRNA-(poly(A) RNA)- Purification Strategy Most Appropriate for Your Situation? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209 How Many Rounds of Oligo(dT)–Cellulose Purification Are Required? . . . . . . . . . . . . . . . . . . . . . . . . . 210 Which Oligo(dT)–Cellulose Format Is Most Appropriate? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210 Can Oligo(dT)–Cellulose Be Regenerated and Reused? . . . 211 Can a Kit Designed to Isolate mRNA Directly from the Biological Sample Purify mRNA from Total RNA? . . . 212 Maximizing the Yield and Quality of an RNA Preparation . . . 212 What Constitutes “RNase-Free Technique”? . . . . . . . . . . . . 212 Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S. Gerstein Copyright © 2001 by Wiley-Liss, Inc. ISBNs: 0-471-37972-7 (Paper); 0-471-22390-5 (Electronic)