Recently Viewed

Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

EcoRI

5'...G↓A A T T C...3'
3'...C T T A A↑G...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Unique buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by overlapping CpG methylation Thermal inactivation at 65°C in 20 min High concentration available Genome qualified Recombinant enzyme LO certified

The EcoRI restriction enzyme recognizes G^AATTC sites and cuts best at 37°C in its own unique buffer
  
Loading...
Lambda DNA digested with EcoRI

Lambda DNA digested with EcoRI, 0.7% agarose, 5 cleavage sites

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsXapI, MunI, TasI.
Conditions for 100% Activity
  • 1X Buffer EcoRI: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, 0.02% Triton X-100, and 0.1 mg/mL BSA
  • Incubate at 37°C
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteLow salt concentration, large excess of the enzyme, pH > 8.0, or the replacement of Mg2+ by Mn2+ may result in star activity.
Storage BufferEcoRI is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 300 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.2 mg/mL BSA, 0.15% Triton X-100, and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)

1X		 G (green)

1X		 O (orange)

1X		 R (red)

1X		 Tango

(yellow)

1X / 2X
EcoRI Buffer [Unique] 37°C 0-20 NR 100 100† NR 100 2X

† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...m5C GAATTm5C G...3'

3'... Gm5CTTAA Gm5C...5'		 Impaired
EcoBI (TGA(N)8TGCT) 5'...TGm6AATTC(N)4 TGCT...3'

3'...AC TTAAG(N)4m6ACGA...5'		 No effect

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
5 0 1 1 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
1 1 1 1 0 1

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition SequenceSecond EnzymeEnzymes recognizing newly generated recognition sequence
G^AATTC
  • MunI (MfeI)/FastDigest MfeI (MunI) (C^AATTG)
  • TasI (Tsp509I)/FastDigest Tsp509I (TasI) (^AATT)
  • TasI (Tsp509I)/FastDigest Tsp509I (TasI)
  • XapI (ApoI)/FastDigest XapI* (R^AATTC)
  • EcoRI/FastDigest EcoRI
  • TasI (Tsp509I)/FastDigest Tsp509I (TasI)
  • XapI (ApoI)/FastDigest XapI
  • XapI (ApoI)/FastDigest XapI* (R^AATTT)
  • TasI (Tsp509I)/FastDigest Tsp509I (TasI)
  • XapI (ApoI)/FastDigest XapI

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition SequenceNewly generated sequence after reactionEnzymes that cleave the newly generated sequence
G^AATTCGAATTAATTC
  • PdmI (XmnI)/FastDigest PdmI
  • FastDigest MseI (SaqAI)
  • [TasI (Tsp509I)/FastDigest Tsp509I (TasI)]
  • Tru1I (MseI)/FastDigest Tru1I
  • VspI (AseI)/FastDigest AseI (VspI)