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R6265

Sigma-Aldrich

EcoR I from Escherichia coli BS5

Restriction Enzyme

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

Quality Level

grade

for molecular biology

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

Certificate of Analysis

Certificate of Origin

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...
Lilia Gabsalilow et al.
Nucleic acids research, 41(7), e83-e83 (2013-02-15)
Targeted genome engineering requires nucleases that introduce a highly specific double-strand break in the genome that is either processed by homology-directed repair in the presence of a homologous repair template or by non-homologous end-joining (NHEJ) that usually results in insertions...
D S Jansson et al.
British poultry science, 53(6), 790-799 (2013-02-13)
1. Vancomycin-resistant Enterococcus faecium (VRE(fm)) has recently spread among Swedish broiler farms. The objectives were to investigate VRE(fm) persistence within barns between flocks, and to determine whether day-old chicks, feed or forklift trucks used for loading crates could be identified...
Diana Schenkwein et al.
Nucleic acids research, 41(5), e61-e61 (2013-01-01)
Integrating viral vectors are efficient gene transfer tools, but their integration patterns have been associated with genotoxicity and oncogenicity. The recent development of highly specific designer nucleases has enabled target DNA modification and site-specific gene insertion at desired genomic loci....

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