Source: Paracoccus carotinifaciens 3K
Unit definition: One unit is defined as the amount of enzyme required to digest a unique site
in 1 μg of DNA pMHgaI/DriI in 1 hour at 37°C in a total reaction volume of 50 μl.
|PcsI activity assay on DNA pMHgaI/DriI
Products were separated in 1,4% agarose gel in Buffer TAE.
Assayed on: pMHgaI/DriI is a linearized plasmid pMHgaI, which carries genes of DNA-methyltransferases M1.HgaI (recognition sequence 5`-GCGTC-3`) and M2.HgaI (5`-GACGC-3`) and includes a unique PcsI canonical site:
Optimal SE-buffer: SE-buffer PcsI
Enzyme activity (%):
Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 0.1 mg/ml BSA, 50% glycerol; Store at -20°C.
Non-specific hydrolisis: No detectable degradation of 1μg of Lambda DNA was observed after incubation with 1 units of enzyme for 16 hours at 37°C in a total reaction volume of 50 μl.Reagents Supplied with Enzyme
10 X SE-buffer PcsI
Methylation sensitivity: The enzyme cleaves only C5-methylated DNA and does not cut unmodified DNA .
Notes: When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
1. Chernukhin V.A., Nayakshina T.N., Tarasova M.V., Golikova L.N., Akishev A.G., Dedkov V.S., Mikhnenkova N.A., Degtyarev S.Kh. Bacterial strain Paracoccus carotinifaciens 3K- producer of PcsI site specific endonuclease. // Russian Federation patent RU 2377294 C1 (2009).