Product SourceAn E. coli strain that carries the cloned BaeI gene from Bacillus sphaericus SDZ-8 (R. Morgan)
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|S-adenosylmethionine (SAM)||-20||32 mM|
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1 µg of Lambda DNA in 1 hour at 25°C in a total reaction volume of 50 µl.
1X CutSmart® Buffer
Supplement with 20 μM S-adenosylmethionine (SAM)
Incubate at 25°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: 100%
NEBuffer 3.1: 50%
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
Heat Inactivation65°C for 20 min
dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Blocked by Some Combinations of Overlapping
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.
- BaeI cleaves DNA substrates twice to excise its recognition site generating a 28 base-pair fragment with 5 base 3´ overhangs.
- BaeI was found to cut at its defined cleavage site as well as one base pair further from its recognition site.
- Requires S-adenosylmethionine for optimal activity (supplied with enzyme).
- Does BaeI have any special requirements?
- Does BaeI cleave DNA twice?
- What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
- Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
- 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?
- Why is my Restriction Enzyme not cutting DNA?