Kro I

5,500.00 22,000.00 

  • CNY: 77.98 ¥ - 311.91 ¥

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

  • Концентрация: 1000 е.а./мл
  • Recognition site: G↑C(5mC)GGC / CGG(5mC)C↓G
  • Источник: Kocurea rosea 307
  • Оптимальный буфер: SE-буфер G
  • Оптимальная температура: 37
  • Температура инактивации, 20 мин при: 65
SKU: E541 Category:

Description

Recognition site:

G↑C(5mC)GGC
CGG(5mC)C↓G

Source: Kocurea rosea 307

Substrate specificity:

Unit definition: One unit is defined as the amount of enzyme required to hydrolyze completely 1 μg of linearized plasmid pMHpaII 1 in 1 hour at 37°C in a total reaction volume of 50 μl.

KroI activity assay on DNA pMHpaII 1/DriI

Lanes:
1 and 6 – 1 Kb SE DNA-markers.
2 – Control DNA pMHpaII 1/DriI,
3 – 0.5 μl Kro I
4 – 1 μl Kro I
5 – 2 μl Kro I

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

KroI activity assay on DNA pMHpaII 1/DriI

Assayed on: DNA pMHpaII 1/DriI is a linearized plasmid pMHpaII 1. pMHpaII 1 carries a gene of DNA-methyltransferase M.HpaII, which methylates sites 5`-CCGG-3` producing 5`-C(5mC)GG-3`/3`-GG(5mC)C-5`, and includes three canonical sites
5`-GC(5mC)GGC-3`/3`-CGG(5mC)CG-5`.
Optimal SE-buffer: SE-buffer G
Enzyme activity (%):

B G O W Y ROSE
50 100 25 50 75 100

Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg /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 30°C in a total reaction volume of 50 μl.Reagents Supplied with Enzyme

10 X SE-buffer G

Methylation sensitivity: The enzyme cleaves C5-methylated DNA and doesn’t cut unmodified DNA [1].

Notes:

References:

1. Dedkov V.S., Rechkunova N.I., Degtyarev S.Kh., Zhilkin P.A., Kolykhalov A.A., Verkhozina V.A. Bacillus sphaericus – a strain producer of restriction endonuclease BsiI.// Soviet Union patent SU 1784642 (1992). (In Russian)
2. Chernukhin V.A., Zhuravleva R.O., Tarasova G.V., Boltengagen A. A., Akishev A.G., Mikhnenkova N.A., Degtyarev S.Kh. Bacterial strain Kocuria rosea – producer of KroI site specific endonuclease. // Russian Federation patent RU 2394099 C1 (2010).

 

Available Options

SKUРackage sizePackagePrice 
E541S50 u5,500.00 
E542L250 u22,000.00