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Restriction Endonuclease

Home > PRODUCTS > Restriction Endonuclease

EZ-CleanCut™ Sac I

No Product Name Cat.# Conc Size Price(₩) Add
1 EZ-CleanCut™ Sac I CR005S 20 units/μl 2,000 units ₩ 68,000 추가
2 EZ-CleanCut™ Sac I CR005M 20 units/μl 4,000 units ₩ 122,400 추가
3 EZ-CleanCut™ Sac I CR005L 20 units/μl 10,000 units ₩ 272,000 추가
4 EZ-CleanCut™ Sac I CR005H 100 units/μl 10,000 units ₩ 272,000 추가
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EZ-CleanCut Sac I with high purity and fidelity has greatly reduced star activity than Sac I. 


Recognition & cleavage sequence



Source: Streptomyces achromogenes

 

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), 100 mM NaCl, 0.1 mM EDTA, 1 mM dithiothreitol, 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 dithiothreitol, 200 μg/ml BSA, 50% glycerol.

 

Heat Inactivation

EZ-CleanCut™ Sac I can be inactivated at 65 for 20 min.

 

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Conditionally sensitive

 

Prolonged incubation

A minimum amount of enzyme required to digest 1-μg substrate DNA for 16 hr; 0.13 U.

 

Relative activity in EzBuffers

EzBuffer I: 75%

EzBuffer II: 50%

EzBuffer III: 10%

EzBuffer IV: 100%

FastCut Buffer: 100%

 

Note

High salts over 50 mM inhibit its activity. Use clean DNA for efficient cleavage. Cleavage of mammalian genomic DNA is blocked by some combinations of overlapping CpG methylation. This shows an impaired result when CG overlap exists like (5’-CGAGCTCG-3'), (5’-CGAGCTC-3’) or (5’-GAGCTCG-3’).

 

"Shipping only to Asia."

● Material Safety Data Sheet

- Enzynomics_MSDS_CR005_EZ-CleanCut Sac 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 hose 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 or 4hr in 50-μl reaction. 

 

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. 

 

Blue/white screening

To test the integrity of DNA ends, a plasmid, pSKM2 containing a unique site in the lacZ alpha gene is digested with 10-fold excess enzyme, ligated, and introduced into DH5α. The transformed cells are plated on X-gal/IPTG/Amp plates. The number of blue and white colonies formed are measured. Blue colonies indicate that an intactness of the test gene is maintained during the cleavage/ligation process. In contrast, white colonies fail to do so. Fewer than 1% white colonies are formed with enzyme.

 

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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