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PluT I

No Product Name Cat.# Conc Size Price(₩) Add
1 PluT I R115S 10 units/μl 500 units ₩ 77,000 추가
2 PluT I R115M 10 units/μl 1,000 units ₩ 138,000 추가
3 PluT I R115L 10 units/μl 2,500 units ₩ 308,000 추가
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Recognition & cleavage sequence


Source: Photorhabdus luminescens

 

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 to digest 1 µg of pBR322 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

 

Storage

10 mM Tris-HCl (pH 7.4), 50 mM KCl, 0.1 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 dithiothreitol, 200 μg/ml BSA, 50% glycerol.

 

Heat Inactivation

PluT I can be inactivated at 65 for 20 min.

 

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Sensitive

 

Relative activity in EzBuffers

EzBuffer I: 75%

EzBuffer II: 25%

EzBuffer III: 10%

EzBuffer IV: 100%

FastCut Buffer: 100%

 

Note

PluT I requires two copies of its recognition sequence for cleavage to occur.

● Material Safety Data Sheet

- Enzynomics_MSDS_R115_PluT 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. 

 

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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