Description
Recognition site and hydrolysis position:
| GGGCCC | GGGCC↑C |
| CCCGGG | C↓CCGGG |
Source: An E.coli strain that carries the cloned Apa I gene from Acetobacter pasteurianus
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-dcm-, BamHI-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA/BamHI
Optimal SE-buffer: SE-buffer Y + BSA
Enzyme activity (%):
| B | G | O | W | Y | ROSE |
| 50 | 25 | 0 | 0 | 100 | 50 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme > 95% 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
For Regular enzyme: 10 X SE-buffer Y, BSA
For Turbo enzyme: 10 X SE-buffer ROSE
Methylation sensitivity: Blocked by overlapping Dcm methylation(CmCWGG): GGGCCCWGGNotes: 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.
References:
Seurinck, J., Van de Voorde, A., Van Montagu, M. Nucleic Acids. Res.11: 4409-4415 (1983).
