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Acc I (Xmi I)

No Product Name Cat.# Conc Size Price(₩) Add
1 Acc I R023S 4 units/µl 1,000 units ₩ 80,000 추가
2 Acc I R023M 4 units/µl 2,000 units ₩ 144,000 추가
3 Acc I R023L 4 units/µl 5,000 units ₩ 320,000 추가
4 Acc I R023H 20 units/µl 5,000 units ₩ 320,000 추가
Product Name Cat.# Size Price(₩) EA Total(₩) 삭제
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Recognition & cleavage sequence


Source: Acinetobacter calcoaceticus


Isoschizomer : Fbl I, Xmi I

 

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 7.5 @ 25), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.15% Triton, 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

Acc I can be inactivated at 80 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: 100%

EzBuffer III: 100%

EzBuffer IV: 100%

FastCut Buffer: 100%

 

Note

It does not cleave DNA with 3 or fewer bases on each side of the recognition site. At least 13 bases are required beyond the ends of the recognition site for efficient cleavage. Cleavage of mammalian genomic DNA is blocked by CpG methylation overlapping its recognition sequence. Both M13 and pUC19 contain a single Acc I site.

● Material Safety Data Sheet

Enzynomics_MSDS_R023_Acc 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.

 

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 digestesd 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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