|Catalog number||E213||E214||E214X |
|Package, u.a.||200||1000||1000 |
|Concentration, u.a./ml||10000||10000||50000 |
|Methylation sensitivity||not tested|
|Origin||E.coli strain that carries the cloned Afe I gene from Alcaligenes faecalis T2774|
|Storage conditions||20 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 1 mM DTT; 200 ug/ml BSA; 50% glycerol; Store at -20°C.|
|Assayed on||Lambda DNA (BamHI-digest)|
|Ligation||After 10-fold overdigestion with enzyme more than 80% of DNA pBR322 fragments can be ligated and recut.|
|Non-specific activity||No nonspecific activity was detected after incubation of 1 ug of DNA with 40 u.a. of enzyme for 16 hours at 37°C.|
|Optimum temperature||37 oC|
|Inactivation 20 minutes||65oC|
|Optimum 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|
Note: The minimum number of units that resulted in complete digestion of 1 ug 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.
References: Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Degtyarev, S.Kh.
Unpublished observations. (1994)