Bmt I

Enzyme nameBmt I
PrototypeNheI
Known analogsAsuNH I, NheI
SKUSE-E457
Turbo versionAvailable
High-concentration versionNot available
Restriction typeType IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… GCTAG▼C …3'
3'… C▲GATCG …5'
SourceAn E.coli strain that carries the cloned Bmt I gene from Bacillus megaterium S2
Optimal bufferSE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
10505010075100
Unit definitionOne 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.
Assayed onLambda DNA (Hind III-digest)
Storage conditions10 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.
LigationAfter 20-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer W
NotesThe 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.
References
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