Recognition site | CATG↑ ↓GTAC |
Source | Flavobacterium aquatile N3 |
Assayed on | pUC19 DNA |
Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of pUC19 DNA in 1 hour at 37°C in a total reaction volume of 50 μl. |
Optimal SE-buffer | SE-buffer FaeI (33 mM Tris-acetate (pH 8.3 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA |
Enzyme activity (%) | B | G | O | W | Y | Rose | 25 - 50 | 50 - 75 | 10 - 25 | 10 - 25 | 75 - 100 | 100 |
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Optimal temperature | 37oC |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
Ligation | After 3-fold overdigestion with enzyme more then 90% of the DNA fragments can be ligated with T4 DNA Ligase at 16°C and recut. |
Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of pUC19 DNA with 2 u.a. of enzyme for 16 hours at 37°C.
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Reagents Supplied with Enzyme |
10 X SE-buffer FaeI, BSA |
Methylation sensitivity | Blocked by CmATG methylation |
Inactivation 20 minutes under | 65oC |
Notes | To obtain 100% activity, BSA should be added the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation. |
Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Chernukhin, V.A., Kileva, E.V., Tomilova, J.E., Dedkov, V.S.. Unpublished observations (2006).
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