OPTIZYME™ EcoRI, Fisher BioReagents™

Descrizione

5'...G^A A T T C...3'
3'...C T T A A^G...5'

Conditions for 100% Activity:

• 1X OPTIZYME Buffer EcoRI: 50mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, 100mM NaCl, 0.02% Triton X-100 and 0.1mg/ml BSA.
• Incubate at 37°C.

Supplied with: 10X OPTIZYME EcoRI Buffer
Enzyme Activity in OPTIZYME buffers:

• Buffer 1: 0 - 20%
• Buffer 2: No reaction
• Buffer 3: 100%
• Buffer 4: No reaction
• Buffer 5: 100%™

™star activity occurs at greater than 5-fold overdigestion (5 units X 1 hour)
Storage Buffer:
10mM potassium phosphate (pH 7.4 at 25°C), 300mM NaCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 0.15% Triton X-100 and 50% (v/v) glycerol.
Ligation and Recleavage:
More than 95% of DNA fragments can be ligated and recut after a 50-fold overdigestion with EcoRI.
Digestion of Agarose-embedded DNA:
A minimum of 5 units of EcoRI is required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
Note: Low salt concentration, large excess of the enzyme, pH>8.0, or the replacement of mg²⁺ by Mn²⁺ may result in star activity.
Compatible Ends: XapI, MunI, TasI.
Methylation Effects:
Dam: never overlaps - no effect.
Dcm: never overlaps - no effect.
CpG: may overlap - cleavage impaired.

• 5'...m5C GAATTm5C G...3'

3'... Gm5CTTAA Gm5...5'

EcoKI: never overlaps - no effect.

EcoBI: may overlap - no effect.

• 5'...TGm6AATTC(N)₄TGCT...3'

3'...AC TTAAG(N)₄m6ACGA...5'