Source: An E.coli strain that carries the cloned Bmt I gene from Bacillus megaterium S2
Assayed on: Lambda DNA (Hind III-digest)
Unit definition: One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (Hind III-digest) in 1 hour at 37°C in a total reaction volume of 50μl.
Optimal SE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Activity in SEBuffers:
SEBuffer B 10 - 25%
SEBuffer G 50 - 75%
SEBuffer O 50 - 75%
SEBuffer W 100%
SEBuffer Y 75 - 100%
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); 200 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol;
Store: at -20°C.
Ligation: After 20-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis: No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme: 10 X SE-buffer W
Methylation sensitivity: not tested
Inactivation: 20 minutes under 65°C
Notes: The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,13. BmtI cleaves linear plasmid DNA at a rate 5 times higher than supercoiled plasmid DNA.
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