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R6265

Sigma-Aldrich

EcoR I from Escherichia coli BS5

Restriction Enzyme

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CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.53

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.

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

pictograms

Health hazard

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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J Hedgpeth et al.
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
C Kessler et al.
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
Jaroslav Jelinek et al.
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
Saqib H Ansari et al.
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
Nico Mitro et al.
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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