| Recognition site | ASST↑
↓TSSA |
| Source | E.coli strain that carries the cloned Set I gene from Streptomyces werraensis 37 |
| Assayed on | Double-stranded oligonucleotide
5`-CGAGTTTATAGCTGGGCCCAAC-3`
3`-GCTCAAATATCGACCCGGGTTG-5` |
| Description | Note! SetI cleaves a canonical site and several other sites with a weaker activity.
In the case of long incubation with SetI DNA can be digested to small oligos. |
| Unit definition | One unit is defined as the amount of enzyme required to cleave 1 pmol of the double-stranded oligonucleotide of the below indicated structure in 1 hour at 55°C in a total reaction volume of
20 μl. |
| Optimal SE-buffer | Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | R | | 25 - 50 | 25 - 50 | 75 - 100 | 75 - 100 | 100 | 100 |
|
| Optimal temperature | 55oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.6); 100 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 50% glycerol; Store at -20°C. |
| Ligations | After 5-fold overdigestion with enzyme, approximately 50% of the pBR322 DNA fragments can be ligated with T4 DNA Ligase and recut. |
| Reagents Supplied with Enzyme | 10 X SE-buffer Y |
| Methylation sensitivity | not tested |
| Inactivation 20 minutes under | 80oC |
| MSDS: | Download MSDS as PDF |
| References: |  Tomilova J.E., Degtyarev S.Kh Substrate specificity of new restriction endonuclease SetI // In press
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