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Key Documents

R6265

Sigma-Aldrich

EcoR I from Escherichia coli BS5

Restriction Enzyme

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About This Item

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

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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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
Cong Zhu et al.
Nucleic acids research, 41(4), 2455-2465 (2013-01-11)
Zinc-finger nucleases (ZFNs) have been used for genome engineering in a wide variety of organisms; however, it remains challenging to design effective ZFNs for many genomic sequences using publicly available zinc-finger modules. This limitation is in part because of potential
Annabel A Ferguson et al.
Methods in molecular biology (Clifton, N.J.), 940, 87-102 (2012-10-30)
The generation of transgenic animals is an essential part of research in Caenorhabditis elegans. One technique for the generation of these animals is biolistic bombardment involving the use of DNA-coated microparticles. To facilitate the identification of transgenic animals within a
TALENs and ZFNs are associated with different mutation signatures.
Yongsub Kim et al.
Nature methods, 10(3), 185-185 (2013-02-12)

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