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Fok I

No Product Name Cat.# Conc Size Price(₩) Add
1 Fok I R088S 5 units/µl 1,000 units ₩ 70,000 추가
2 Fok I R088M 5 units/µl 2,000 units ₩ 126,000 추가
3 Fok I R088L 5 units/µl 5,000 units ₩ 280,000 추가
Product Name Cat.# Size Price(₩) EA Total(₩) 삭제
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Recognition & cleavage sequence


Source: Flavobacterium okeanokoites

 

Reaction conditions

1X EzBuffer IV, 37

1X FastCut Buffer, 37

 

FastCut Buffer

Enzynomics restriction enzyme can cut substrate DNA in 5-15 min with FastCut Buffer

 

1X EzBuffer IV

20 mM Tris-acetate (pH 7.9 @ 25), 50 mM potassium acetate, 10 mM magnesium acetate, 100 μg/ml BSA

 

Unit definition

One unit is defined as the amount of enzyme required for complete digestion of 1-μg bacteriophage λ at 37 for 1 hr in 50-μl reaction mixtures.

 

Storage

10 mM Tris-HCl(pH 7.4 @ 25), 50 mM NaCl, 1 mM dithiothreitol, 0.1 mM EDTA, 200 μg/ml BSA, 0.1% tween 20, 50% Glycerol.

 

Dilution buffer: EzDiluent A

10 mM Tris-HCl (pH 7.4 @ 25), 50 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA, 50% glycerol.

 

Heat Inactivation

Fok I can be inactivated at 65 for 20 min.

 

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Conditionally sensitive

CpG methylation: Conditionally sensitive

 

Prolonged incubation

A minimum amount of enzyme required to digest 1-μg substrate DNA for 16 hr; 1 U.

 

Relative activity in EzBuffers

EzBuffer I: 100%

EzBuffer II: 100%

EzBuffer III: 10%

EzBuffer IV: 100%

FastCut Buffer: 100%

 

Note

Cleavage is inhibited by dcm methylation and CpG methylation partially overlapping its cleavage site.

Overdigestions of >5 units of FokI per μg of DNA and incubation times >2 hours are not recommended.

● Material Safety Data Sheet

- Enzynomics_MSDS_R088_Fok I

 

● Quality control

Overdigestion assay

DNA digested for 16 hr in 50-μl reaction with 100 U of enzyme resulted in same the DNA band patterns as those obtained with 1 U of enzyme for 1 hr.

 

Endonuclease assay

Less than 5% of 1-μg of ΦX174 RFI is converted to RFII when the DNA is incubated with 50 U of enzyme at 37 for 4hr in 50-μl reaction. 


Ligation and recutting

More than 95% of DNA fragments (1 μg) digested with 20-fold excess enzyme can be ligated by T4 DNA ligase (400 U) at 16 for 16 hr. Of the ligation products, > 95% can be re-cut. 


Extreme pure (EP)

No detectable degradation of 32P-end labeled single-stranded and double-stranded (5’-, 3’-overhang and blunt end) oligonucleotides occurred during incubation with 100 U of enzyme for 4 hr.

 

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