Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S Gerstein opyright◎2001 ISBNS:0-471-37972-7( Paper);0-47 (Electronic) 4 Nucleic Acid Hybridization Sibylle Herzer and david F Englert Planning a Hybridization Experiment The Importance of Patience 40l What Are your most essential needs? Is a More Sensitive Detection System Always Better?..401 Visualize Your Particular Hybridization Event 403 What Can You conclude from commercial Sensitivity Data? Labeling issues 403 Which Labeling Strategy ls Most Appropriate for Your situation? 403 What Criteria Could You Consider When Selecting Label? 405 Radioactive and Nonradioactive Labeling Strategies Compared What Are the Criteria for Considering direct over Indirect Nonradioactive Labeling Strategies?........... 410 What Is the st storage Stability of Labeled Probes? Should the Probe Previously Used within the Hybridization Solution of an Earlier Experiment Be Applied in a New Experiment? ...4|2 How Should a probe be denatured for reuse? Is It Essential to Determine the Incorporation Efficiency of Every Labeling Reaction? Is It Necessary to Purify Every Probe ..413399 14 Nucleic Acid Hybridization Sibylle Herzer and David F. Englert Planning a Hybridization Experiment . . . . . . . . . . . . . . . . . . . . . 401 The Importance of Patience . . . . . . . . . . . . . . . . . . . . . . . . . . 401 What Are Your Most Essential Needs? . . . . . . . . . . . . . . . . . 401 Visualize Your Particular Hybridization Event . . . . . . . . . . . . 401 Is a More Sensitive Detection System Always Better? . . . . . 403 What Can You Conclude from Commercial Sensitivity Data? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 403 Labeling Issues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 403 Which Labeling Strategy Is Most Appropriate for Your Situation? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 403 What Criteria Could You Consider When Selecting a Label? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 405 Radioactive and Nonradioactive Labeling Strategies Compared . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 409 What Are the Criteria for Considering Direct over Indirect Nonradioactive Labeling Strategies? . . . . . . . . . . . 410 What Is the Storage Stability of Labeled Probes? . . . . . . . . . 411 Should the Probe Previously Used within the Hybridization Solution of an Earlier Experiment Be Applied in a New Experiment? . . . . . . . . . . . . . . . . . . . . . . 412 How Should a Probe Be Denatured for Reuse? . . . . . . . . . . 412 Is It Essential to Determine the Incorporation Efficiency of Every Labeling Reaction? . . . . . . . . . . . . . . . . . . . . . . . . . 412 Is It Necessary to Purify Every Probe? . . . . . . . . . . . . . . . . . 413 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)