| Recognition site | ↑CATG
GTAC↓ |
| Source | An E.coli strain that carries the cloned Fat I gene from Flavobacterium aquatile NL3 |
| 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 55°C in a total reaction volume of 50 μl. |
| Optimal SE-buffer | G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 10 - 25 | 100 | 25 - 50 | 10 - 25 | 50 - 75 | 100 |
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| Optimal temperature | 55oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; and 50% glycerol. Store at -20°C. |
| Ligation | After 2-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 DNA with 3 u.a. of enzyme for 16 hours at 55°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer G |
| Methylation sensitivity | Blocked by mCATG methylation |
| Inactivation 20 minutes under | 65oC |
| Notes | The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,25.
FatI cleaves linear plasmid DNA at a rate 1.5-2 times higher than supercoiled plasmid DNA.
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| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: | Dedkov, V.S., Kileva, E.V., Popichenko, D.V., Degtyarev, S.K. Biotekhnologia #5, 3-7 (2002)
 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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