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Ava II (Eco47 I)

No Product Name Cat.# Conc Size Price(₩) Add
1 Ava II R040S 10 units/µl 2,000 units ₩ 80,000 추가
2 Ava II R040M 10 units/µl 4,000 units ₩ 144,000 추가
3 Ava II R040L 10 units/µl 10,000 units ₩ 320,000 추가
4 Ava II R040H 50 units/µl 10,000 units ₩ 320,000 추가
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Recognition & cleavage sequence


Source: Anabaena variabilis

 

IsoschizomerAfl I, Bme18 I, Eco47 I, Sin 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 7.4 @ 25), 50 mM KCl, 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

Ava II can be inactivated at 80 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; 0.25 U.


Relative activity in EzBuffers

EzBuffer I: 100%

EzBuffer II: 100%

EzBuffer III: 50%

EzBuffer IV: 100%

FastCut Buffer: 100%

 

Note

Activity is not affected by supercoiled DNA structure. Cleavage of mammalian genomic DNA is blocked by CpG methylation overlapping its recognition sequence. It is markedly affected by impurities in DNA.

 

● Material Safety Data Sheet

- Enzynomics_MSDS_R040_Ava II

 

● 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. 

 

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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