Recently Viewed

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

EcoO109I (DraII)

5'...R G↓G N C C Y...3'
3'...Y C C N G↑G R...5'
Available as a FastDigest enzyme for rapid DNA digestion

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

The EcoO109I (DraII) restriction enzyme recognizes RG^GNCCY sites and cuts best at 37°C in Tango buffer (Isoschizomers: DraII)
  
Loading...
Lambda DNA digested with EcoO109I (DraII)

Lambda DNA digested with EcoO109I (DraII), 0.7% agarose, 3 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

EcoO109I is blocked by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer Tango: 33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, and 0.1 mg/mL BSA
  • Incubate at 37°C
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme is 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.
Storage BufferEcoO109I is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.2 mg/mL BSA, 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
Tango 37°C 50-100 20-50 20-50 20-50 100 100 1X or 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: may overlap - blocked.
  • CpG: may overlap - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.
Methylation typeSequenceCleavage effect
Dcm (CCWGG)5'...RGGNCm5CT GG...3'
3'...YCCNG GAm5CC...5' 		Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
12345
50-100

Number of recognition sites in DNA molecules

LambdaΦX174M13mp18/19pBR322puc18/19pUC57
300411
pTZ19R/UpTZ57RpBluescriptIIKS(-/+)pBluescriptIISK(-/+)pACYC177pACYC184
001114

New sites generated by ligation

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

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
RG^GCCCYRGGCCGCCCY
  • [BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)]
  • [CviJI]
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • TauI/FastDigest TauI
RG^GGCCYRGGGCGGCCY
  • [BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)]
  • [CviJI]
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • TauI/FastDigest TauI