R6265
EcoR I from Escherichia coli BS5
Restriction Enzyme
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About This Item
CAS Number:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.53
Recommended Products
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.
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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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)
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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
Yu-Feng Huang et al.
BMC systems biology, 6 Suppl 2, S10-S10 (2013-01-11)
Current next-generation sequencing (NGS) platforms adopt two types of sequencing mechanisms: by synthesis or by ligation. The former is employed by 454 and Solexa systems, while the latter by SOLiD system. Although the pros and cons for each sequencing mechanism
Rachel M Smith et al.
Nucleic acids research, 41(1), 391-404 (2012-11-14)
Type IIB restriction-modification systems, such as BcgI, feature a single protein with both endonuclease and methyltransferase activities. Type IIB nucleases require two recognition sites and cut both strands on both sides of their unmodified sites. BcgI cuts all eight target
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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