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EcoO109I (DraII)

Kód produktu: ER0261 Kód výrobce: ER0261 Kód dodavatele: {AAF88562-5451-442C-BF3C-4D5EC5305F96} Výrobce: Life Technologies Czech Republic s.r.o.
2 571,25 Kč
2 125,00 Kč bez DPH
do týdne
2000 units

5'...R GG N C C Y...3'
3'...Y C C N GG R...5'

Zobraz detailní popis
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Detailní popis

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.

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 DNA Minimum 5 units of the enzyme is required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double Digestion Perform double digestion using DoubleDigest.
Hazardous No
Isoschizomers Search for commercial isoschizomers using REsearch.
Storage Buffer EcoO109I 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.
Storage Condition -20 C

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 type Sequence Cleavage 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
1 2 3 4 5
50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
3 0 0 4 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
0 0 1 1 1 4

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^GCCCY RGGCCGCCCY
  • [BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)]
  • [CviJI]
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • TauI/FastDigest TauI
RG^GGCCY RGGGCGGCCY
  • [BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)]
  • [CviJI]
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • TauI/FastDigest TauI

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