Source: Bacillus sphaericus
5000-10000 u/ml Store at -20°C
Recognition Sequence:
↑(N)7GAAGNNNNNNTAC(N)12↑
↓(N)12CTTCNNNNNNATG(N)7↓
Source: Bacillus sphaericus
Supplied in: 20mM KH2PO4 (pH 7.4); 100mM KCl; 0.1mM EDTA; 200 µg/ml BSA,7mM 2-mercaptoethanol; 50% glycerol.
Reaction Conditions:
1X SEBuffer 2K
Incubate at 37°C.
Warranty period for the enzyme storage at -20˚C is two years from the date of the last assay indicated on the enzyme vial.
1X SEBuffer 2K (pH 7.6 @ 25ºC)
10mM Tris-HCl, 200mM KCl, 10mM MgCl2, 1 mM DTT
Unit Definition: One unit is defined as the amount of enzyme required to digest 1 µg of T7 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.
Quality Control Assays
Ligation: After 3-fold overdigestion with Bar I, ~90% of T7 DNA fragments can be ligated with T4 DNA Ligase and ~95% of these can be recut.
16-Hour Incubation: A 50 ml reaction containing 1µg of T7 DNA and 4 units of enzyme incubated for 16 hours resulted in the same pattern of DNA bands as a reaction incubated for 1 hour.
Oligonucleotide Assay: No detectable degradation of a single- and double-stranded oligonucleotide was observed after incubation with 2 units of enzyme for 3 hours.
Enzyme Properties
Activity in SEBuffers:
SEBuffer B 0%
SEBuffer G 0-10%
SEBuffer O 25-50%
SEBuffer W 50-75%
SEBuffer Y 10-25%
SEBuffer ROSE 40%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Heat Inactivation: Yes (65°C for 20 minutes)
Reagents supplied with enzyme: 10X SEBuffer 2K
| Product | Description |
| E547 | Bar I 100 Units |
| E548 | Bar I 500 Units |
Datasheet
Stability Test Report
Material Safety Datasheet
