The enzyme cleaves only C5-methylated DNA and does not cut unmodified DNA.
10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA; 100 µg/ml BSA; 7 mM 2-mercaptoethanol; 50% glycerol.
1X SEBuffer PcsI
Incubate at 37°C.
1X SEBuffer PcsI (pH 8.3 @ 25ºC)
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 site 5`-A(5mC)GNNNNNNN(5mC)GT-3` in 1 μg of DNA pMHgaI/DriI in 1 hour at 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 recognition sequence.
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