| Recognition site | GAT↑C
C↓TAG |
| Source | Bacillus stearothermophilus KT |
| Assayed on | Lambda DNA (dam-) |
| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) 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.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 25 - 50 | 50 - 75 | 75 - 100 | 100 | 50 - 75 | 100 |
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| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C. |
| Ligations | After 5-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 5 u.a. of enzyme for 16 hours at 37°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer W |
| Methylation sensitivity | Blocked by overlapping Dam methylation (GmATC) GATC
.
Not blocked by CG methylation.
Cut hemimethylated site:
5’- GmATC-3’ / 5’-GATC-3’ |
| Inactivation 20 minutes under | 65oC |
| MSDS: | Download MSDS as PDF |
| References: | Nadeev A.N., Chernukhin V. A., Sevastyanova O.O., Tomilova J.E., Shinkarenko N.M., Evdokimov A.A., Degtyarev S. Kh. BstKTI, a New dam-Sensitive Neoschizomer of Restriction Endonuclease MboI, which is Able to Cleave Hemimethylated Substrate.// Biotekhnologia (Moscow), No.2, 5-10 (2006). (In Russian)
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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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