Recognition site | AGC↑GCT TCG↓CGA |
Source | An E.coli strain that carries the cloned Afe I gene from Alcaligenes faecalis T2774 |
Assayed on | Lambda DNA (BamHI-digest) |
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. |
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 | Rose | 10 - 25 | 25 - 50 | 75 - 100 | 75 - 100 | 100 | 100 |
|
Optimal temperature | 37oC |
Storage conditions | 10 mM Tris-HCl (pH 7.6); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
Ligation | After 10-fold overdigestion with enzyme more than 80% of DNA pBR322 fragments can be ligated and recut. |
Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of DNA with 40 u.a. of enzyme for 16 hours at 37°C.
|
Reagents Supplied with Enzyme |
10 X SE-buffer Y
|
Methylation sensitivity | not tested |
Inactivation 20 minutes under | 65oC |
Notes | The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,25.
AfeI cleaves supercoiled and linear plasmid DNA (pBR322) at a roughly equal rate. AfeI cleaves Lambda DNA/BamHI digest at a rate 3-4 times higher than plasmid DNA.
|
Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Degtyarev, S.Kh.
Unpublished observations. (1994)
|