Now comes supplied with 1 vial of Gel Loading Dye, Purple (6X), (B7024 )
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cloned at neb recombinant timesaver 5min icon_CutSmart dil_A-grey incubation temp heat inactivation
Catalog #SizeConcentrationPriceQtyAdd to Cart
R0108S3,000 units10,000 units/ml$61.00Add to Cart
R0108T3,000 units50,000 units/ml$61.00Add to Cart
R0108L15,000 units10,000 units/ml$246.00Add to Cart
R0108M15,000 units50,000 units/ml$246.00Add to Cart
Restriction Endonucleases: H-M,
Time-Saver™ Qualified Restriction Enzymes
Restriction Enzyme Digestion


HaeIII crystals (Sandra Cook, Lydia Dorner and Ira Schildkraut, New England Biolabs)

HaeIII crystals (Sandra Cook, Lydia Dorner and Ira Schildkraut, New England Biolabs)

Product Source

An E. coli strain that carries the HaeIII gene from Haemophilus aegypticus (ATCC 11116).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
CutSmart® Buffer-2010X

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X CutSmart® Buffer
Incubate at 37°C

1X CutSmart® Buffer:
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 μg/ml BSA
pH 7.9 @ 25°C

Activity in NEBuffers

NEBuffer 1.1: 50%
NEBuffer 2.1: 100%
NEBuffer 3.1: 25%
CutSmart® Buffer: 100%

Diluent Compatibility

Storage Temperature


Storage Conditions

10 mM Tris-HCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
50 mM KCl
pH 7.4 @ 25°C

Heat Inactivation

80°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive


  1. GGCm5C is cleaved, but GGm5CC is resistant to cleavage.


  1. Is HaeIII affected by methylation?
  2. Does HaeIII cleave single-stranded DNA?
  3. Is HaeIII used in special techniques?
  4. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  5. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  6. How can I access the old NEBuffer™ Activity Chart?
  7. 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?
  8. Has NEB used any enzymes in Chromatin Conformation Capture techniques, such as 3C or HiC? 
  9. Why is my Restriction Enzyme not cutting DNA?
  10. Why do I see a DNA smear on an agarose gel after a restriction digest?
  11. Why do I see additional DNA bands on my gel after a restriction digest?
  12. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  13. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?

Tech Tips

Restriction Enzymes in Chromatin Conformation Capture


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

Selection Charts

Usage Guidelines & Tips

Troubleshooting Guides

Interactive Tools

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
  • Ligation and Recutting (Terminal Integrity):
    After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.
  • Non-Specific DNase Activity (16 hour, Buffer):
    The buffer is tested in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]


The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.