Aox I

6,000.00 24,000.00 

  • CNY: 465.36 ¥ - 1,861.44 ¥

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

  • Концентрация: 500 е.а./мл
  • Recognition site: ↑RG(5mC)Y / ↓RG(5mC)Y
  • Источник: Arthrobacter oxydans 25K
  • Оптимальный буфер: SE-буфер AoxI
  • Оптимальная температура: 60
  • Температура инактивации, 20 мин при: N

Available Options

SKUРackagePackagePrice 
E569S50 u6,000.00 
  • CNY: 465.36 ¥
E570L250 u24,000.00 
  • CNY: 1,861.44 ¥

Description

Recognition site and hydrolysis position:

RG(5mC)Y ↑RG(5mC)Y
Y(5mC)GR Y(5mC)GR↓

Source: Arthrobacter oxydans 25K
Substrate specificity:
Unit definition: One unit is defined as the amount of enzyme
required to hydrolyze in 1 μg of linearized plasmid pMHaeIII/DriI in 1 hour at 60°C in a total reaction volume of 50 μl. As a result a linearized plasmid pMHaeIII/DriI disappears (see run 4 in the figure).

AoxI activity assay on DNA pMHaeIII/DriI
Lanes:
1 – Control DNA pMHaeIII/DriI,
2 – 0.5 μl Aox I
3 – 1 μl Aox I
4 – 2 μl Aox I
5 – 1 Kb SE DNA-markers

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

AoxI activity assay on DNA pMHaeIII/DriI

Assayed on: DNA pMHaeIII/DriI is a linearized plasmid pMHaeIII. pMHaeIII carries a gene of DNA-methyltransferase M.HaeIII, which methylates sites 5`-GGCC-3` producing 5`-GG(5mC)C-3`/3`-C(5mC)GG-5`.
Optimal SE-buffer: SE-buffer AoxI
Enzyme activity (%):

B G O W Y ROSE
75 25 10 25 75 100

Storage conditions: 10 mM Tris-HCl (pH 7.4); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethan. 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 60°C in a total reaction volume of 50 μl. Reagents Supplied with Enzyme 10 X SE-buffer AoxI
Methylation sensitivity: The enzyme cleaves C5-methylated DNA and doesn’t cut unmodified DNA.
Notes: References:
1. Chernukhin V.A., Belichenko O.A., Tarasova M.V., Gonchar D.A., Akishev A.G., Dedkov V.S., Mikhnenkova N.A., Degtyarev S.Kh. Bacterial strain Arthrobacter oxydans – producer of AoxI site specific endonuclease. // Russian Federation patent RU 2399663 C1 (2009).