Source: An E.coli strain that carries recombinant plasmids.
Assayed on: Lambda DNA
Unit definition: One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Optimal SE-buffer K (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM KCl; 1 mM DTT.)
Activity in SEBuffers:
SEBuffer B 10 - 25%
SEBuffer G 25 - 50%
SEBuffer O 50 - 75%
SEBuffer W 50 - 75%
SEBuffer Y 25 - 50%
SEBuffer ROSE 100%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Optimal temperature: 37°C
Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 50% glycerol.
Store: at -20°C
Ligation: After 5-fold overdigestion with enzyme 80% of the DNA fragments can be ligated. Of these 90% can be recut. In the presence of 10% PEG ligation is better.
Non-specific hydrolisis: No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 1 hours at 37°C.
Reagents Supplied with Enzyme: 10 X SE-buffer K
Methylation sensitivity: not tested
Inactivation: 20 minutes under 80°C
Notes: High enzyme concentration may result in star activity or incomplete DNA cleavage. We recommend increasing incubation time instead of using an excess of Bpu10I. The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,5.
Bpu10 I - CC↑TNAGC - GGANT↓CG - Restriction Endnuclease from Sibenzyme
* availibility of sample size may be limited
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