Product SourceAn E. coli strain that carries the AlwI gene from Acinetobacter lwoffii (R. Morgan).
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (dam-) in 1 hour at 37°C in total reaction volume of 50 µl.
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 NEBuffersNEBuffer 1.1: 50%
NEBuffer 2.1: 50%
NEBuffer 3.1: 10%
CutSmart® Buffer: 100%
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
pH 7.4 @ 25°C
dam methylation: Blocked
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Quality Control AssaysThe 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.
- 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):
The product is tested for non-specific nuclease degradation 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.
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please contact NEB’s Global Business Development team at firstname.lastname@example.org.
- Is AlwI blocked by dam methylation?
- Why is an extended digest not recommended?
- Why is AlwI- cut DNA difficult to ligate?
- How can I increase ligation efficiency?
- What is the activity of AlwI at 25°C?
- My restriction enzyme used to be available at a lower concentration. Why does it now come at a higher concentration of 10,000 u/ml?
- My enzyme is no longer Time-Saver™ qualified. What happened?
- What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
- How can I access the old NEBuffer Activity Chart?
- 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?
- My restriction enzyme used to work well in the old NEBuffer but the new Performance chart indicates it has lower activity even though the only difference is the addition of BSA and removal of DTT to the new buffers. Why?
- Why is my Restriction Enzyme not cutting DNA?
- Why do I see a DNA smear on an agarose gel after a restriction digest?
- Why do I see additional DNA bands on my gel after a restriction digest?
- How many nucletotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
- Alphabetized List of Recognition Specificities
- Compatible Cohesive Ends and Generation of New Restriction Sites
- Cross Index of Recognition Sequences
- Dam-Dcm and CpG Methylation
- Enzymes with Nonpalindromic Sequences
- Frequencies of Restriction Sites
- Time-Saver™ Qualified Enzymes
- Type IIS Restriction Enzymes
- Why Choose Recombinant Enzymes?
Usage Guidelines & Tips
- Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
- Activity of Restriction Enzymes in a Q5®, Taq or Phusion PCR Mix
- Cleavage Close to the End of DNA Fragments
- Dam and Dcm Methylases of E. coli
- Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
- Double Digests
- Effects of CpG Methylation on Restriction Enzyme Cleavage
- Heat Inactivation
- Megabase Mapping
- 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
This enzyme produces a single base extension, which is difficult to ligate.