Recognition Sequence :
GCTAG↑C
C↓GATCG
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.
Reaction Temperature :
37°C
Storage Buffer :
10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol
Ligation :
After 20-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and recut.
Source :
An E.coli strain that carries the cloned Bmt I gene from Bacillus megaterium S2
Assayed on :
Lambda DNA (Hind III-digest)
Working buffer :
W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
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.
Concentration, u.a./ml :
20000
Inactivation :
20min Under 65°C
Reagents Supplied :
10 X SE-buffer W
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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