|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 (%)|
|10 - 25||25 - 50||75 - 100||75 - 100||100||100|
|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Unpublished observations.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)|
|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