Recognition Sequence :
GATNN↑NNATC
CTANN↓NNTAG
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.
Reaction Temperature :
60°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 about 80% of the DNA fragments can be ligated and recut.
Source :
Bacillus species 8
Working buffer :
G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Non-specific hydrolisis :
Long incubation is not recommended. Extended incubations and high concentration of the enzyme may result in star activity.
Concentration, u.a./ml :
5000
Inactivation :
20min Under 80°C
Methylation sensitivity :
Blocked by overlapping Dam-methylation (GmATC): 5’-GmATCN^NNATC-3’/3’-CTmAGN^NNTAG.
Blocked by overlapping (5mC)-methylation: 5’-GATNN^NNAT(5mC)-3’/3’-(5mC)TANN^NNTAG-5’.
Cuts hemimethylated sequence: 5’-GATNN^NNAT(5mC)-3’/3’-CTANN^NNTAG-5’.
Reagents Supplied :
10 X SE-buffer G
Notes :
High enzyme concentration (more than 5-fold overdigestion on 1 μg substrate) may result in star activity.
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