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.
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 50 μl. |
Recognition sites | GCTAG↑C |
Source | E.coli strain that carries the cloned Bmt I gene from Bacillus megaterium S2 |
Assayed on | Lambda DNA (Hind III-digest) |
Optimal buffer | W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) |
Optimal temperature | 37°C |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
Ligations | 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. |
Methlation effect | Methylation sensitivity not tested |
Thermal inactivation | enzyme is inactivated by incubation at 65°C for 20 mins. |
Campatible Ends | AsuNHI?^, BspOI, NheI^ |
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