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EcoR I

No Product Name Cat.# Conc Size Price(₩) Add
1 EcoR I R002S 20 units/µl 20,000 units ₩ 57,000 추가
2 EcoR I R002M 20 units/µl 40,000 units ₩ 102,600 추가
3 EcoR I R002L 20 units/µl 100,000 units ₩ 228,000 추가
4 EcoR I R002H 100 units/µl 100,000 units ₩ 228,000 추가
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Recognition & cleavage sequence


Source: E. coli RY13

 

Reaction conditions

1X EzBuffer EcoR I, 37

2X FastCut Buffer, 37

 

FastCut Buffer

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

 

1X EzBuffer EcoR I

100 mM Tris-HCl (pH 7.5 @ 25), 50 mM NaCl, 10 mM MgCl2, 0.025% Triton X-100

 

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 K-PO4 (pH 7.5 @ 25), 300 mM NaCl, 0.1 mM EDTA, 10 mM 2-mercaptoethanol, 0.15% Triton X-100, 200 μg/ml BSA, 50% glycerol.

 

Dilution buffer: EzDiluent C

10 mM Tris-HCl (pH 7.4 @ 25), 250 mM NaCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.15% Triton X-100, 200 μg/ml BSA, 50% glycerol.

 

Heat Inactivation

EcoR 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: 50%

EzBuffer II: 100%

EzBuffer III: 75%

EzBuffer IV: 100%

FastCut Buffer: 100%

 

Note

It can cleave non-specifically the sequences of AATT or PuPuATPyPy (Pu=purine; Py=pyrimidine) (star activity). The addition of 2 mM β-mercaptoethanol may inhibit this star activity. Long-term storage can result in aggregation, leading to reduced activity. Both DNA and RNA in a DNA/RNA hybrid duplex can be cleaved sequence specifically. Cleavage of mammalian genomic DNA is impaired by overlapping CpG methylation. Reaction condition of low salt, excess enzyme, excess glycerol (>5%) or high pH (>8.0) can result in star activity.

● Material Safety Data Sheet

- Enzynomics_MSDS_R002_EcoR 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.

 

● Related paper

1. Hyo Young Park, et al (2012). PLOS-one 7, e45812

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