Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S Gerstein opyright◎2001 ISBNS:0-471-37972-7( Paper);0-471 (Electronic) PCR Kazuko Aoyag Introduction 292 Developing a PCR Strategy: The Project Stage 293 Assess Your needs Identify Any Weak Links in Your PCR Strategy 295 Manipulate the Reaction to Meet Your Needs Developing a PCr Strategy: The Experimental Stage 296 What Are the Practical Criteria for Evaluating a DNA Polymerase for Use in PCR! 296 How Can Nucleotides and Primers Affect a PCr Reaction How Do the Components of a Typical PCR Reaction Buffer Affect the reaction? How Can You Minimize the Frequency of Template What makes for good pos and Negative Amplification Controls? What makes for a reliable control for gene Expression? 309 ifferent Cycling Parameters Af PCR Reaction? Instrumentation: By What Criteria Could You Evaluate a Thermocycler? 309 How Can Sample Preparation Affect Your results?11 PCR Kazuko Aoyagi Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292 Developing a PCR Strategy: The Project Stage . . . . . . . . . . . . . 293 Assess Your Needs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 293 Identify Any Weak Links in Your PCR Strategy . . . . . . . . . . 295 Manipulate the Reaction to Meet Your Needs . . . . . . . . . . 296 Developing a PCR Strategy: The Experimental Stage . . . . . . . 296 What Are the Practical Criteria for Evaluating a DNA Polymerase for Use in PCR? . . . . . . . . . . . . . . . . . . . . . . . . 296 How Can Nucleotides and Primers Affect a PCR Reaction? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303 How Do the Components of a Typical PCR Reaction Buffer Affect the Reaction? . . . . . . . . . . . . . . . . . . . . . . . . . 305 How Can You Minimize the Frequency of Template Contamination? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 306 What Makes for Good Positive and Negative Amplification Controls? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 308 What Makes for A Reliable Control for Gene Expression? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 309 How Do the Different Cycling Parameters Affect a PCR Reaction? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 309 Instrumentation: By What Criteria Could You Evaluate a Thermocycler? . . . . . . . . . . . . . . . . . . . . . . . . . . 309 How Can Sample Preparation Affect Your Results? . . . . . . . 311 291 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)