Product Class: Restriction Endonuclease

HindIII-HF®
neb31 cloned at NEB recombinant engineered time saver dil_B 37 80 Heat

HindIII-HF has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10119499. 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^AGCT_T
Isoschizomers | Single Letter Code

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/AGCTT
Catalog # Size Concentration
R3104S 10000.0 units 20000 units/ml
R3104T 10000.0 units 100000 units/ml
R3104L 50000.0 units 20000 units/ml
R3104M 50000.0 units 100000 units/ml

Featured Videos

View Video Library
  • ReduceStarActivity_video_thumb

    Reduce Star Activity with High-Fidelity Restriction Enzymes

  • timesaverprotocol_video_thumb

    TIME-SAVER™ Protocol for Restriction Enzyme Digests

  • NEBTVEp15VLthumb280x210

    NEB® TV Ep. 15 – Applications of Restriction Enzymes

  • cuttingclose_video_thumb

    Restriction Enzyme Digest Protocol: Cutting Close to DNA End

  • dnasmear_video_thumb

    Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel

  • EnzymeNotCutting_video_thumb

    Why is My Restriction Enzyme Not Cutting DNA?

  • TooManyBands_video_thumb

    Restriction Enzyme Digest Problem: Too Many DNA Bands

  • nebcloner_dd_demo_thumb

    Double Digestion with NEBcloner

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  3. Restriction Digest Protocol
  4. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  5. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. When should I choose the HF version of an enzyme?
  2. When is star activity a concern?
  3. Why does the HF version of the enzyme have a different recommended buffer than the wild type enzyme?
  4. When should I choose the High Fidelity (HF®) version of the enzyme?
  5. How is the improvement in fidelity of HF restriction endonucleases quantitated?
  6. What is the Fidelity Index (FI)?
  7. Can the change in buffer preference of the HF enzyme be advantageous?
  8. Will the HF enzyme produce elevated star activity when used in a buffer other than the one recommended?
  9. What does it mean to be Time-Saver™ qualified?
  10. What does HF® refer to following the name of a restriction enzyme?
  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. 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?
  14. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
  15. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  16. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  17. Does NEB offer any BSA-free and/or animal origin-free restriction enzymes for linearization of plasmids for mRNA vaccine development?
  18. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Troubleshooting

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