Recognition Sequence :
GAT↑C
C↓TAG
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) 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); 100 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol.
Ligation :
After 5-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Source :
Bacillus stearothermophilus KT
Assayed on :
Lambda DNA (dam-)
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 5 u.a. of enzyme for 16 hours at 37°C.
Concentration, u.a./ml :
2000-5000
Inactivation :
20min Under 65°C
Methylation sensitivity :
Blocked by overlapping Dam methylation (GmATC) GATC .
Not blocked by CG methylation.
Cut hemimethylated site: 5`- GmATC-3` / 5`-GATC-3`
Reagents Supplied :
10 X SE-buffer W
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