| Recognition site | GCATC(N)5↑
CGTAG(N)9↓ |
| Source | An E.coli strain that carries the cloned SfaN I gene from Streptococcus faecalis N |
| 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 | O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 10 - 25 | 25 - 50 | 100 | 75 - 100 | 0 - 10 | 25 |
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| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 300 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
| Ligation | After 10-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer O
For E165T and E166T an additional 10X SE-buffer ROSE is supplied. |
| Methylation sensitivity | not tested |
| Inactivation 20 minutes under | 80oC |
| Notes | Turbo SfaN I can be used for short time (10 min) DNA digestion as well as for standard reaction. The reaction can be performed using optimal or universal (ROSE) Buffer
Turbo DNA Digestion:
Applications:
-Fast DNA analysis
-Fast preparation of vectors for cloning
-Double digestion
Enzyme Properties:
1 μl of Turbo SfaN I cuts 1 μg of DNA in 1 x SE-Buffer O or universal 1 x SE-Buffer ROSE in 10 min (see the protocol below). A short time of DNA digestion requires high quality purification of DNA sample. The enzyme may be used in standard conditions of DNA cleavage (E165).
Turbo reaction protocol:
20 μl of the reaction volume:
10 x SE Buffer ROSE - 2 μl
DNA - 0,2-1 μg
Nuclease-free water - to 20 μl
+ 1μl of Turbo SfaN I
Incubate at 37°C for 10 min.
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| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: | Schildkraut, I., Greenough, L. Unpublished observations.
 V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.2, No 3, pp 39-46, 2006
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Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:
To view the fragments length values please point mouse cursor over diagram |
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322
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