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Cfr42 I (Sac II)

No Product Name Cat.# Conc Size Price(₩) Add
1 Cfr42 I R080S 10 units/μl 1,200 units ₩ 63,000 추가
2 Cfr42 I R080M 10 units/μl 2,400 units ₩ 113,400 추가
3 Cfr42 I R080L 10 units/μl 6,000 units ₩ 252,000 추가
4 Cfr42 I R080H 50 units/μl 6,000 units ₩ 252,000 추가
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Recognition & cleavage sequence


Source: Citrobacter freundii RFL42


Isoschizomer : Ksp I, Sac II, Sfr303 I, SgrB I, Sst II

 

Reaction conditions

1X EzBuffer I, 37

1X FastCut Buffer, 37

 

FastCut Buffer

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

 

1X EzBuffer I

10 mM Bis Tris Propane-HCl (pH 7.0 @ 25), 10 mM MgCl2, 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 KPO4 (pH 7.4 @ 25), 100 mM NaCl, 0.1 mM EDTA, 7 mM 2-mercaptoethanol, 200 /ml BSA, 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

Cfr42 I can be inactivated at 65 for 20 min.

 

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: sensitive

 

Prolonged incubation

A minimum amount of enzyme required to digest 1-μg substrate DNA for 16 hr; 0.5 U.

 

Relative activity in EzBuffers

EzBuffer I: 100%

EzBuffer II: 50%

EzBuffer III: 25%

EzBuffer IV: 75%

FastCut Buffer: 100%

 

Note

Cleavage of mammalian genomic DNA is blocked by CpG methylation.

● Material Safety Data Sheet

- Enzynomics_MSDS_R080_Cfr42 I

 

● Quality control

Overdigestion assay

DNA digested for 16 hr in 50-μl reaction with 20 U of enzyme resulted in same the DNA band patterns as those obtained with 1 U of enzyme for 1 hr.

 

Ligation and recutting

More than 95% of DNA fragments (1 μg) digested with 50-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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