Product info: Afe I


Name
Afe I
Cat. #E213E214E214X
Package, u.a.20010001000
Concentration, u.a./ml100001000050000

Recognition site
AGCGCT
TCGCGA
SourceE.coli strain that carries the cloned Afe I gene from Alcaligenes faecalis T2774
Assayed onLambda DNA (BamHI-digest)
Unit definitionOne 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-bufferY (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Enzyme activity (%)
BGOWYRose
10 - 2525 - 5075 - 10075 - 100100100
Optimal temperature
37oC
Storage conditions10 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.
LigationsAfter 10-fold overdigestion with enzyme more than 80% of DNA pBR322 fragments can be ligated and recut.
Non-specific hydrolisisNo 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 sensitivitynot tested
Inactivation 20 minutes under
65oC
NotesThe 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 Unpublished observations.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
Fragment lengths
M12345M

M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322