Unpublished data
Unpublished observations (1994).
PspOM I
| Enzyme name | PspOM I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | (ApaI) | ||||||||||||
| Known analogs | Bsp120I, Apa I | ||||||||||||
| SKU | SE-E215 | ||||||||||||
| Turbo version | Not available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Restriction type | Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites. | ||||||||||||
| Recognition site | 5'… G▼GGCCC …3' 3'… CCCGG▲G …5' | ||||||||||||
| Source | An E.coli strain that carries the cloned PspOM I gene from Pseudomonas species OM2164 | ||||||||||||
| Optimal buffer | SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) | ||||||||||||
| Optimal temperature | 37 °C | ||||||||||||
| Inactivation temperature | 65 °C | ||||||||||||
| Buffer activity |
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| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (BamHI-digest) in 1 hour at 37°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | Lambda DNA (BamHI-digest) | ||||||||||||
| Storage conditions | 20 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; and 50% glycerol. Store at -20°C. | ||||||||||||
| Ligation | After 10-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut. | ||||||||||||
| Nonspecific hydrolysis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C. | ||||||||||||
| Methylation sensitivity | not tested | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer Y | ||||||||||||
| Notes | The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,13. | ||||||||||||
| References |