Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S Gerstein ISBNS:0-471-37972-7( Paper);0-471 (Electronic) 9 Restriction endonucleases Derek Robinson, Paul R. Walsh, and joseph A. Bonventre Background Information 226 Which Restriction Enzymes Are Commercially Why Are Some Enzymes More Expensive Than Others? 227 What Can You do to reduce the cost of Working with Restriction Enzymes? If You Could Select among Several Restriction Enzymes for Your Application, What Criteria Should You Consider to Make the Most Appropriate Choice? 229 What Are the General Properties of Restriction Endonucleases? 232 What Insight Is Provided by a Restriction Enzyme's Quality Control data? .233 How Stable Are Restriction Enzymes How Stable Are Diluted Restriction Enzymes 236 Simple Digests How Should You Set up a Simple Restriction Digest?... 236 ls It Wise to Modify the Suggested Reaction Conditions? 237 Complex Restriction Dige Should You Alter the Reaction Volume and DNAst...239 How Can a Substrate Affect the restriction Dige Concentration? ...24 Double Digests: Simultaneous or Sequential? 225225 9 Restriction Endonucleases Derek Robinson, Paul R. Walsh, and Joseph A. Bonventre Background Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226 Which Restriction Enzymes Are Commercially Available? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226 Why Are Some Enzymes More Expensive Than Others? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227 What Can You Do to Reduce the Cost of Working with Restriction Enzymes? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228 If You Could Select among Several Restriction Enzymes for Your Application, What Criteria Should You Consider to Make the Most Appropriate Choice? . . . . . . . . . . . . . . 229 What Are the General Properties of Restriction Endonucleases? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232 What Insight Is Provided by a Restriction Enzyme’s Quality Control Data? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233 How Stable Are Restriction Enzymes? . . . . . . . . . . . . . . . . . . 236 How Stable Are Diluted Restriction Enzymes? . . . . . . . . . . . 236 Simple Digests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236 How Should You Set up a Simple Restriction Digest? . . . . . 236 Is It Wise to Modify the Suggested Reaction Conditions? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237 Complex Restriction Digestions . . . . . . . . . . . . . . . . . . . . . . . . . 239 How Can a Substrate Affect the Restriction Digest? . . . . . 239 Should You Alter the Reaction Volume and DNA Concentration? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241 Double Digests: Simultaneous or Sequential? . . . . . . . . . . . . 242 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)