10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA;
2-mercaptoethanol; 200 µg/ml
1XSEBuffer W. Incubate at 55°C.
W (pH 8.5 @ 25ºC)
10 mM Tris-HCl 150 mM
10 mM MgCl2; 1 mM
Unit Definition: One unit is defined as the amount of enzyme required
to hydrolyze completely1 μg of linearized plasmid pHspAI10/DriI+M.Fsp4HI
in 1 hour at 55°C in a total reaction volume of 50 μl.
pHspAI10/DriI+M.Fsp4HI is a plasmid pHspAI10, which is linearized with
DriI, and, additionally, modified with Fsp4HI DNA methyltransferase
pHspAI10 carries a gene of HspAI DNA methyltransferase, that modifies the
sequence . 5`-GCGC-3`, producing 5`-G(5mC)GC-3`.
M.Fsp4HI modifies the sequence 5`-GCNGC-3`, producing 5`-G(5mC)NGC-3`.
A substrate pHspAI10/DriI+M.Fsp4HI includes one site
which is MteI canonical site . The enzyme activity depends on a number and
positions of methylated nucleotides in the recognition sequence.
For example, MteI cuts the recognition site with six 5-methylcytosines, but the
enzyme activity is reduced for more that one order 
Quality Control Assays
No detectable degradation of 1μg of Lambda DNA was observed after
incubation with 10 units of enzyme for 16 hours at 55°C
in a total reaction
volume of 50 μl.
No detectable degradation of a single- and double-stranded oligonucleotide
was observed after incubation with 10 units of enzyme for