Pcs I

5,500.00 22,000.00 

  • CNY: 67.50 ¥ - 269.99 ¥

MD DNA endonuclease Pcs I.
The enzyme cleaves C5-methylated DNA and doesn’t cut unmodified DNA.

  • Концентрация: 1000 е.а./мл
  • Recognition site: (5mC)GNNNNN↑NN(5mC)G / G(5mC)NN↓NNNNNG(5mC)
  • Источник: Paracoccus carotinifaciens 3K
  • Оптимальный буфер: SE-буфер PcsI
  • Оптимальная температура: 37
  • Температура инактивации, 20 мин при: 65
SKU: E505 Category:

Description

Recognition site:

(5mC)GNNNNN↑NN(5mC)G
G(5mC)NN↓NNNNNG(5mC)

Source: Paracoccus carotinifaciens 3K

Substrate specificity:

Unit definition: 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.

PcsI activity assay on DNA pMHgaI/DriI

Lanes:
1 and 6 – 1 Kb SE DNA-markers
2 – Control pMHgaI/DriI
3 – 0.5 μl Pcs I
4 – 1 μl Pcs I
5 – 2 μl Pcs I

Products were separated in 1,4% agarose gel in Buffer TAE.

PcsI activity assay on DNA pMHgaI/DriI

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:
5`-W(5mC)GNNNNNNN(5mC)GW-3`/
3`-WG(5mC)NNNNNNNG(5mC)W-5` [1].
Optimal SE-buffer: SE-buffer PcsI
Enzyme activity (%):

B G O W Y ROSE
50 25 0 10 50 20

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.

Ligation: –

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 [1].

Notes: When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion.

References:

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).

 

Available Options

SKUРackage sizePackagePrice 
E505S50 u5,500.00 
E506L250 u22,000.00