1000 u/ml store at -20°C
Source: Paracoccus carotinifaciens 3K
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.
Optimal recognition site (100% activity ):
Quality Control Assays
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 3 hours.
Activity in SEBuffers:
SEBuffer B 50-75%
SEBuffer G 25-50%
SEBuffer O 0%
SEBuffer W 10-25%
SEBuffer Y 50-75%
SEBuffer ROSE 20%
When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Reagents supplied with enzyme:
10X SEBuffer PcsI
Yes (65°C for 20 minutes)
Restriction Endnuclease from Sibenzyme
* availibility of sample size may be limited
Final price excl. shipping costs3