| Recognition site | C↑TCGAG
GAGCT↓C |
| Source | Streptomyces fradiae 274 |
| Assayed on | Lambda DNA (Hind III-digest) |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (Hind III-digest) in 1 hour at 50°C in a total reaction volume of 50 μl. |
| Optimal SE-buffer | B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 100 | 75 - 100 | 50 - 75 | 50 - 75 | 75 - 100 | 100 |
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| Optimal temperature | 50oC |
| 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. |
| Ligations | After 20-fold overdigestion with enzyme more than 90% 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 50°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer B |
| Methylation sensitivity | Blocked by CTCGmAG methylation.
Not blocked by CTmCGAG methylation. |
| Inactivation 20 minutes under | 65oC |
| Notes | At 37°C activity is about 70% from maximum. |
| MSDS: | Download MSDS as PDF |
| References: | Puchkova, L.I., Krivopalova, G.N., Andreeva, I.S., Selina, A.V., Serov, G.D., Rechkunova, N.I., Degtyarev, S.Kh. Izv. Sib. Otd. Akad. Nauk SSSR 1: 32-34 (1990).
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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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