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SibEnzyme Ltd. :: Display product information: Afe I
Afe I
Catalog numberE213E214E214X
Package, u.a.20010001000
Concentration, u.a./ml100001000050000

Recognition siteAGC^GCT
Methylation sensitivitynot tested
OriginE.coli strain that carries the cloned Afe I gene from Alcaligenes faecalis T2774
Storage conditions20 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 1 mM DTT; 200 ug/ml BSA; 50% glycerol; Store at -20C.
Assayed onLambda DNA (BamHI-digest)
LigationAfter 10-fold overdigestion with enzyme more than 80% of DNA pBR322 fragments can be ligated and recut.
Non-specific activityNo nonspecific activity was detected after incubation of 1 ug of DNA with 40 u.a. of enzyme for 16 hours at 37C.
Optimum temperature37 oC
Inactivation 20 minutes65oC
Optimum SE-bufferY (33 mM Tris-acetate (pH 7.9 at 25C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)

Enzyme % activity:
10 - 2525 - 5075 - 10075 - 100100

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 Unpublished observations.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)