Recognition Sequence :
GGCCNNNN↑NGGCC
CCGGN↓NNNNCCGG
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 50°C in a total reaction volume of 50 μl.
Reaction Temperature :
50°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 10-fold overdigestion with enzyme more than 70% of the DNA fragments can be ligated and recut. In the presence of 10% PEG ligation is better.
Source :
An E.coli strain, that carries the cloned gene Sfi I from Streptomyces fimbriatus
Working buffer :
G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 20 u.a. of enzyme for 16 hours at 50°C.
Concentration, u.a./ml :
10000
Inactivation :
20min Under 65°C
Methylation sensitivity :
Blocked by overlapping Dcm methylation (CmCWGG): GGCCWGGNNGGCC.
Not blocked by overlapping Dcm methylation (CmCWGG): GGCCNNNNNGGCCWGG.
Reagents Supplied :
10 X SE-buffer G, BSA
Notes :
Incubation at 37°C rusults in 75-100% activity.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
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