Protocols

1. Optimizing Restriction Endonuclease Reactions
2. Restriction Digest Protocol
3. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

• Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
• Activity of Restriction Enzymes in PCR Buffers
• Cleavage Close to the End of DNA Fragments
• Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
• Double Digests
• Heat Inactivation
• NEBuffer Activity/Performance Chart with Restriction Enzymes
• Optimizing Restriction Endonuclease Reactions
• Restriction Endonucleases - Survival in a Reaction
• Restriction Enzyme Diluent Buffer Compatibility
• Restriction Enzyme Tips
• Single Letter Codes
• Star Activity
• Traditional Cloning Quick Guide

Selection Charts

• Alphabetized List of Recognition Sequences
• Cleavage of Supercoiled DNA
• Compatible Cohesive Ends and Generation of New Restriction Sites
• Dam-Dcm and CpG Methylation
• Frequencies of Restriction Sites
• Isoelectric Points (pI) for Restriction Enzymes
• Isoschizomers
• NEB Diluent and Buffer Table
• Time-Saver™ Qualified Enzymes
• Why Choose Recombinant Enzymes?

Web Tools

• Competitor Cross-Reference Tool
• DNA Sequences and Maps Tool

• Double Digest Finder
• Enzyme Finder
• NEBcutter™ v3.0
• NEBioCalculator®
• REBASE®

FAQs

1. How can I search for a restriction enzyme by sequence, overhang or name?
2. How should I stop my restriction digest?
3. How stable is a particular restriction enzyme?
4. When is star activity a concern?
5. How should I set up a restriction digest?
6. I don't see any cleavage after my restriction digest. What factors can interfere with cleavage?
7. How can I generate a restriction enzyme site map for my sequence?
8. What information is available in the Restriction Enzyme Database (REBASE)?
9. Do degenerate recognition sites need to be palindromic?
10. Is extended digestion (incubation times > 1 hour) recommended?
11. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
12. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
13. What does it mean to be Time-Saver™ qualified?
14. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
15. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Troubleshooting

• Restriction Enzyme Troubleshooting Guide

Tech Tips

Citations & Technical Literature