Recognition site | ↑(N)8AAGNNNNNCTT(N)13↑ ↓(N)13TTCNNNNNGAA(N)8↓ |
Source | Flavobacterium aquatile Ob10 |
Assayed on | Lambda DNA |
Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl. |
Optimal SE-buffer | W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + SAM |
Enzyme activity (%) | B | G | O | W | Y | Rose | 0 - 10 | 25 - 50 | 75 - 100 | 100 | 50 - 75 | 70 |
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Optimal temperature | 37oC |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 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 20% of the DNA fragments can be ligated and of these 80% can be recut. In the presence of 10%PEG ligation is better. |
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.
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Reagents Supplied with Enzyme |
10 X SE-buffer W, SAM |
Methylation sensitivity | not tested |
Inactivation 20 minutes under | 65oC |
Notes | High enzyme concentration results in star activity
To obtain 100% activity, SAM should be added to a final concentration 0.01 mM
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Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Dedkov, V.S., Sinichkina, S.A., Abdurashitov, M.A., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2001).
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