The Anza Restriction Enzyme Cloning System—a complete, one-buffer, one-protocol system for beautifully simple cloning. Learn more

EcoRI

Catalog number: 15202039

Invitrogen™  Related applications: Restriction Enzyme Cloning

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Description

5´-G↓AATTC-3´ 3´-CTTAA↑G-5´

Invitrogen conventional restriction enzymes come with Universal Buffers L, M, H, K, or T (+BSA). Full details for the this restriction enzymes characteristics are included in the tables below.

Consider upgrading to the Invitrogen™ Anza™ restriction enzyme cloning system for superior convenience. Anza restriction enzymes utilize a single buffer and protocol—complete digestion in 15 minutes at 37°C for all types of DNA substrates. Click here for the Anza equivalent restriction enzyme.
Concentration50 U/ µL
ContentsEcoR I 10X Buffer H (1mL) 10X Loading Buffer (1mL)
Reaction Temperature37°C
Reaction Mixture EcoR I 1 µL 10X Buffer H 2 µL Substrate DNA < 1 µg Sterile water to 20 µL
Heat inactivationEnzyme inactivated by heating at 65°C for 20 minutes.
Star ActivityUnrelated site may be cut in the presence of high concentrations of glycerol or Mn2+, and at low ionic strength. Enzyme activity reduces to 70-80% by the addition of spermine (0.2mM), but star activity reduces to 30-50%.
Effect of DNA MethylationThe sequence GAATTCG is affected by CG methylase.
Unit DefinitionOne unit is the amount of enzyme required to completely digest 1 µg of λDNA in 50 µL of the reaction mixture in one hour at 37°C.
10X Buffer H 500 mM Tris-HCl, pH 7.5 100 mM MgCl2 10 mM Dithiothreitol 1,000 mM NaCl
SourceEscherichia coli RY 13

Relative Activity in Universal Reaction Buffers

BufferLMHKT(+BSA)
Relative Activity (%)20*100*100120*80*
* weak star activity is detected.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Enzyme: EcoR I
Product Size: 20,000 units
Concentration: 30-60 U⁄µl
Star Activity: May exhibit star activity
Compatible Buffer: 10X Buffer H
Methylation Sensitivity: CpG methylation-sensitive
Optimal Reaction Temperature: 37° C
Sensitive to Heat Inactivation: Yes (Heat-Inactivatable)
Shipping Condition: Approved for shipment on Wet or Dry Ice

Contents & storage

Storage Buffer
10 mM Tris-HCl, pH 7.5
100 mM KCl
0.1 mM EDTA
1 mM Dithiothreitol (DTT)
0.15% Triton® X-100
0.01% BSA
50% (v⁄v) glycerol

Loading Buffer
1% SDS
50% glycerol
0.05% Bromophenol blue

Store enzyme and buffer at -20°C, and loading buffer at RT.

Documents

Manuals & protocols

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