Product Class: Restriction Endonuclease

SpeI-HF®
neb31 cloned at NEB recombinant engineered time saver dil_C 37 80 Heat

SpeI-HF has been reformulated, and now also includes Recombinant Albumin (rAlbumin), beginning with Lot #10192521. Learn more.

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

NEB high fidelity restriction endonuclease that recognizes the sequence A^CTAGT

Product Introduction

High-Fidelity (HF®) restriction enzymes have the same specificity as native enzymes, but have been engineered for significantly reduced star activity and performance in a single buffer (rCutSmart Buffer).  All HF-restriction enzymes come with Gel Loading Dye, Purple (6X).  Enjoy the enhanced performance and added value of our engineered enzymes at the same price as the native enzyme:

  • Engineered for improved performance
  • 100% activity in rCutSmart Buffer
  • Time-Saver™ qualified for digestion in 5-15 minutes
  • Reduced star activity
  • Supplied with 1 vial of Gel Loading Dye, Purple (6X)
  • Restriction Enzyme Cut Site: A/CTAGT
Catalog # Size Concentration
R3133S 500.0 units 20000 units/ml
R3133L 2500.0 units 20000 units/ml
R3133M 2500.0 units 100000 units/ml

Protocols, Manuals & Usage

Protocols

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

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

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. What does HF® refer to following the name of a restriction enzyme?
  5. When is star activity a concern?
  6. How should I set up a restriction digest?
  7. I don't see any cleavage after my restriction digest. What factors can interfere with cleavage?
  8. How can I generate a restriction enzyme site map for my sequence?
  9. What information is available in the Restriction Enzyme Database (REBASE)?
  10. Do degenerate recognition sites need to be palindromic?
  11. Is extended digestion (incubation times > 1 hour) recommended?
  12. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  13. 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?
  14. Is Gel Loading Dye, Purple (6X) or Gel Loading Dye, Purple (6X), no SDS compatible with other DNA binding dyes such as SYBR® and GelRed™ during gel electrophoresis?
  15. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
  16. What does it mean to be Time-Saver™ qualified?
  17. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  18. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  19. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Troubleshooting