| Recognition site | GACGC(N)5↑
CTGCG(N)10↓ |
| Source | An E.coli strain that carries the cloned HgaI gene from Haemophilus gallinarum |
| Assayed on | DNA pBR322 |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of DNA pBR322 in 1 hour at 37°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 | Rose | | 100 | 75 - 100 | 10 - 25 | 25 - 50 | 50 - 75 | 50 |
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| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
| Ligation | After 3-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut. |
| Non-specific hydrolisis | Incubation with > 2 units of HgaI per 1 μg of DNA and digestion > 1 hour are not recommended.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer B |
| Methylation sensitivity | Blocked by CG methylation. |
| Inactivation 20 minutes under | 65oC |
| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: | Brown N.L., Smith M. Proc. Natl. Acad. Sci. U. S. A. 74: 3213-3216 (1977).
Sugisaki H.
Gene 3: 17-28 (1978).
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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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