Recognition Sequence :
RG↑GWCCY
YCCWG↓GR
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA/HindIII 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; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Ligation :
After 5-fold overdigestion with enzyme more than 70% of the DNA fragments can be ligated and more than 80% of these can be recut.
Source :
Pseudomonas species PP
Assayed on :
Lambda DNA/HindIII
Working buffer :
Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 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 Dcm methylation(CmCWGG): RGGWCCTGG.
Reagents Supplied :
10 X SE-buffer Y, BSA
Notes :
To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
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