Pcs I

6,000.00 24,000.00 

  • CNY: 479.22 ¥ - 1,916.88 ¥

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

  • Концентрация: 5000 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

Available Options

SKUРackagePackagePrice 
E505S200 u6,000.00 
  • CNY: 479.22 ¥
E506L1000 u24,000.00 
  • CNY: 1,916.88 ¥

Description

Recognition site and hydrolysis position:

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

 
Source: An E.coli strain that carries the cloned Pcs I gene from Paracoccus carotinifaciens 3K.
Substrate specificity:
Unit definition: 
One unit is defined as the amount of enzyme required to digest 1 µg of pMHpySE49/DriI DNA in 1 hour at 37°C in a total reaction volume of 50 µl. As a result, a linearized plasmid pMHpySE49/DriI disappears, and two DNA fragments are produced.
 

PcsI activity assay on pMHpySE49/DriI DNA
Lanes:1 and 7 – 1 Kb SE DNA-ladders
2 – Control pMHpySE49/DriI
3 – 0.25 μl Pcs I
4 – 0.5 μl Pcs I
5 – 1 μl Pcs I
6 – 2 μl Pcs I
Products were separated in 1,4% TAE agarose gel.

Assayed on:
pMHpySE49 plasmid DNA, linearized with DriI (pMHpySE49/DriI).
pMHpySE49 (3536 bp) was obtained by a ligation of the vector pMTL22/FauNDI/SmaI and a PCR-fragment containing M.HpySE526I (forms 5′-A(5mC)GT-3′) gene and
DNA sequence:
5′-GACGTATAAAACGTT-3′
3′-CTGCATATTTTGCAA-5′
Thus, the plasmid has the optimal site of PcsI:
5′-A(5mC)GNNNNNNN(5mC)GT-3′
3′-TG(5mC)NNNNNNNG(5mC)A-5′
Optimal SE-buffer: SE-buffer PcsI
Enzyme activity (%):

B G O W Y ROSE
50-75 50-75 0 10-25 50-75 50

 
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).