10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA; 100 µg/ml BSA;
7 mM 2-mercaptoethanol; 50% glycerol.
Incubate at 37°C.
1X SEBuffer PcsI (pH 8.3 @
10 mM Tris-HCl 20 mM NaCl
3 mM MgCl2 1 mM DTT
One unit is defined as the amount of enzyme required to digest a unique
5`-A(5mC)GNNNNNNN(5mC)GT-3` in 1 μg of DNA pMHgaI/DriI in 1 hour
37°C in a total reaction volume of 50 μl.
DNA pMHgaI/DriI is a linearized plasmid pMHgaI, which included a genes
of DNA-methyltransferases M1.HgaI (recognition sequence 5’-GCGTC-3’)
and M2.HgaI (5’-GACGC-3’) and contains a unique PcsI canonical site:
5'-W(5mC)GNNNNNNN(5mC)GW-3'/3'-WG(5 m C)NNNNNNNG(5mC)W-5'.
The enzyme activity depends on number and position of methylated nucleotides in the
Optimal recognition site (100% activity ):
Incubation: No detectable degradation of 1 µg of λ DNA was
observed after incubation with 1 units of enzyme for 16 hours at 37ºC
No detectable degradation of a single- and double-stranded oligonucleotide
was observed after incubation with 1 units of enzyme for