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Kpn2 I (BspE I)

No Product Name Cat.# Conc Size Price(₩) Add
1 Kpn2 I R110S 10 units/μl 500 units ₩ 73,000 추가
2 Kpn2 I R110M 10 units/μl 1,000 units ₩ 131,000 추가
3 Kpn2 I R110L 10 units/μl 2,500 units ₩ 292,000 추가
4 Kpn2 I R110H 50 units/μl 2,500 units ₩ 292,000 추가
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Recognition & cleavage sequence


Source: Klebsiella pneumoniae RFL2


Isoschizomer : Acc III, Aor13H I , Blf I , BseA I, BsiM I, Bsp13 I, BspE I, BspM II, Mro I

 

Reaction conditions

1X EzBuffer I, 55

1X FastCut Buffer, 55

 

FastCut Buffer

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

 

1X EzBuffer I

10 mM Bis Tris propane-HCl (pH 7.0 @ 25), 10 mM MgCl2, 100 μg/ml BSA

 

Unit definition

One unit is defined as the amount of enzyme required to digest 1 μg of λ DNA in 1 hour at 55°C in a total reaction volume of 50 μl.

 

Storage

10 mM Tris-HCl (pH 7.4), 100 mM KCl, 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

Kpn2 I can be inactivated at 80 for 20 min.

 

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Not sensitive

CpG methylation: Sensitive

 

Relative activity in EzBuffers

EzBuffer I: 100%

EzBuffer II: 25%

EzBuffer III: 75%

EzBuffer IV: 50%

FastCut Buffer: 100%

 

Note

It is an isoschizomer of Acc III and BspE I. Cleavage of mammalian genomic DNA is blocked by CpG methylation.

● Material Safety Data Sheet

- Enzynomics_MSDS_R110_Kpn2 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.

 

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 4 hr 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 lac Z 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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