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Cfr10 I

No Product Name Cat.# Conc Size Price(₩) Add
1 Cfr10 I R114S 10 units/μl 200 units ₩ 85,000 추가
2 Cfr10 I R114M 10 units/μl 400 units ₩ 153,000 추가
3 Cfr10 I R114L 10 units/μl 1,000 units ₩ 340,000 추가
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Recognition & cleavage sequence


Source: Citrobacter freundii RFL 10

 

Reaction conditions

1X EzBuffer Cfr10 I, 37

1X FastCut Buffer, 37

 

FastCut Buffer

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

 

1X EzBuffer Cfr10 I

10 mM Tris-HCl (pH 8.5 @ 25), 3 mM MgSO4, 100 mM KCl, 0.02% Triton X-100

 

Unit definition

One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

 

Storage

10 mM KPO4 (pH 7.4), 200 mM NaCl, 1 mM EDTA, 1 mM DTT, 200 μg/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 DTT, 200 μg/ml BSA, 50% glycerol.

 

Heat Inactivation

No

 

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Sensitive

 

Relative activity in EzBuffers

EzBuffer I: 10%

EzBuffer II: 10%

EzBuffer III: 10%

EzBuffer IV: 25%

FastCut Buffer: 100%

 

Note

Reaction condition with excess enzyme (10 fold) or low salt concentration may result in star activity. For cleavage with Cfr10 I at least two copies of its recognition sequence are required. It is an isoschizomer of BsrF I.

● Material Safety Data Sheet

- Enzynomics_MSDS_R114_Cfr10 I

 

● Quality control

Overdigestion assay

No detectable change in the specific fragmentation pattern is observed after a 5-fold overdigestion with Cfr10 I.

 

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 5-fold excess enzyme can be ligated by T4 DNA ligase (400 U) at 16 for 16 hr. Of the ligation products, ~90% 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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