| Recognition site | AT↑CGAT
TAGC↓TA |
| Source | Bacillus stearothermophilus 29 |
| 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 | G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 25 - 50 | 100 | 50 - 75 | 50 - 75 | 75 - 100 | 100 |
|
| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C. |
| Ligation | 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 40 u.a. of enzyme for 16 hours at 37°C.
|
| Reagents Supplied with Enzyme |
10 X SE-buffer G, BSA (except E205T and E206T).
|
| Methylation sensitivity | Blocked by overlapping Dam methylation (GmATC): GATCGATC.
Blocked by CG methylation. |
| Inactivation 20 minutes under | 65oC |
| Notes | To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
|
| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: | Repin, V.E., Degtyarev, S.K.
Unpublished observations.
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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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