R6265
EcoR I from Escherichia coli BS5
Restriction Enzyme
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grade
for molecular biology
Quality Level
form
buffered aqueous glycerol solution
concentration
10,000 units/mL
shipped in
wet ice
storage temp.
−20°C
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Specificity
Recognition sequence: 5′-G/AATTC-3′
Cutting results: a 2-10-fold EcoR I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: 65 °C for 20 minutes.
Cutting results: a 2-10-fold EcoR I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: 65 °C for 20 minutes.
Application
EcoRI is a restriction endonuclease that is used in molecular biology applications to cleave DNA at the recognition site 5′-G/AATTC-3′, generating fragments with 5′-cohesive termini.
Other Notes
Supplied with 10x Restriction Enzyme Buffer SH (B3657).
Comment: Avoid suboptimal reaction conditions such as low salt, high pH (>8.0) and high glycerol (>5%) as they will alter EcoRI specificity and precipitate star activity.
Unit Definition
One unit is the enzyme activity that completely cleaves 1 mg of λDNA in 1 hour at 37 °C in a total volume of 50 ml of 1x digestion buffer SH for restriction enzymes. 1 mg pBR322 DNA is digested completely by 2 units of EcoR I.
Physical form
Solution in 10 mM Tris-HCl, pH 7.2 , 1 mM EDTA, 200 mM NaCl , 0.5 mM dithioerythritol, 50% glycerol (v/v), 0.2% Triton X- 100 (v/v), 4°C
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Resp. Sens. 1
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WGK 3
flash_point_f
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flash_point_c
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Proceedings of the National Academy of Sciences of the United States of America, 69(11), 3448-3452 (1972-11-01)
The sequence of DNA base pairs adjacent to the phosphodiester bonds cleaved by the RI restriction endonuclease in unmodified DNA from coliphage lambda has been determined. The 5'-terminal nucleotide labeled with (32)P and oligonucleotides up to the heptamer were analyzed
Gene, 92(1-2), 1-248 (1990-08-16)
The properties and sources of all known class-I, class-II and class-III restriction endonucleases (ENases) and DNA modification methyltransferases (MTases) are listed and newly subclassified according to their sequence specificity. In addition, the enzymes are distinguished in a novel manner according
Epigenetics, 7(12), 1368-1378 (2012-10-19)
Genome wide analysis of DNA methylation provides important information in a variety of diseases, including cancer. Here, we describe a simple method, Digital Restriction Enzyme Analysis of Methylation (DREAM), based on next generation sequencing analysis of methylation-specific signatures created by
Journal of pediatric hematology/oncology, 35(4), e153-e156 (2013-02-08)
β-thalassemia is characterized by impaired β-chain synthesis leading to ineffective erythropoiesis, severe anemia, and a need for blood transfusion. Presence of Xmn I polymorphism (-158 C-T nucleotide change) in γ-globin gene is associated with a higher fetal hemoglobin and a
Methods in molecular biology (Clifton, N.J.), 952, 137-144 (2012-10-27)
The role of certain amino acids in the interactions of ligands with their cognate nuclear receptors is usually achieved by the resolution of the crystal structure of the receptor complexed with the ligand. As a complementary functional approach, site-directed mutagenesis
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