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Restriction Endonucleases >
Restriction Endonucleases >
BanII |
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 Recognition Site:


 isoschizomers | compatible ends | single letter code
Source: An E. coli strain that carries the cloned BanII gene from Bacillus aneurinolyticus (IAM 1077)
Reagents Supplied: NEBuffer 4
Enzyme Properties

 Activity in NEBuffers:
| NEBuffer 1: |  | 100% | | NEBuffer 2: |  | 10% | | NEBuffer 3: |  | 50% | | NEBuffer 4: |  | 100% |
When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Methylation Sensitivity:
| dam methylation: Not sensitive | | dcm methylation: Not sensitive | | CpG methylation: Not sensitive | |
More information about: Methylation Sensitivity |
Heat Inactivation: 80°C for 20 minutes
Survival in a Reaction: (+) Intermediate activity. Suitable for extended digestion, but < 8 hours. More information about: Extended Digests with Restriction Enzymes
Reaction & Storage Conditions

 Reaction Conditions: 1X NEBuffer 4 Incubate at
37°C.
1X NEBuffer 4: 20 mM Tris-acetate 50 mM potassium acetate 10 mM Magnesium Acetate 1 mM Dithiothreitol
pH 7.9 @ 25°C
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.
Concentration: 10,000 units/ml
Unit Assay Substrate: λ DNA
Storage Conditions: 10 mM Tris-HCl 50 mM NaCl 1 mM Dithiothreitol 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol
pH 7.4 @ 25°C
Storage Temperature: -20°C
Diluent Compatibility: Diluent A
FAQs


- Do degenerate recognition sites need to be palindromic?
Quality Control for Current Lot

 Quality control values for a specific lot can be found on the datacard which accompanies each product.
Ligation and Re-cutting:
After a 10-fold overdigestion with BanII, > 95%
of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM)
at 16ºC. Of these ligated fragments, > 95% can be recut with BanII.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA
and 50 units of BanII incubated for 16 hours at 37ºC
resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. However, fragments produced by noncanonical cleavage due to star activity may be observed with 10 units of enzyme in similar conditions.
Exonuclease Activity:
Incubation of a 50 μl reaction containing 50 units of BanII with 1 μg of a mixture of single and double-stranded
[3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC
released < 0.3% of the total radioactivity.
Reagents Sold Separately

 NEBuffer 4
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New
England Biolabs, Inc. is an ISO 9001 and ISO
14001 Certified Company.
NEB certifies that it is a small business in accordance with the US Small Business Administration and 13 CFR 121.201 |
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