Recognition Sequence :
GGCCGG↑CC
CC↓GGCCGG
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Ad2 DNA 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 KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.
Ligation :
After 3-fold overdigestion with enzyme > 95% of Ad2 DNA fragments can be ligated with T4 DNA Ligase and recut.
Source :
Rhizobium yangligense
Assayed on :
Adenovirus -2 DNA
Working buffer :
SE-buffer RigI (10 mM Tris-HCl (pH 8.5 at 25°C); 5 mM MgCl2; 1mM DTT.) + BSA
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of Ad2 DNA with 4 units of enzyme for 16 hours at 37°C.
Concentration, u.a./ml :
2000
Inactivation :
20min Under 65°C
Methylation sensitivity :
Blocked by mCG or GmC methylation:
5`-GGC(m5C)GGCC-3`/3`-CCGG(m5C)CGG-5` or
5`-GG(m5C)CGG(m5C)C-3`/3`-C(m5C)GGC(m5C)GG-5`
Reagents Supplied :
10 X SE-buffer RigI, BSA
Storage :
-20°C
Storage at -70°C is recommended for periods longer than 7 days.
Notes :
Do not use BSA for long incubation.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
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