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.
Enzyme Activity in OPTIZYME buffers:
- Buffer 1: 0 - 20%
- Buffer 2: No reaction
- Buffer 3: 100%
- Buffer 4: No reaction
- Buffer 5: 100%™
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.
Dam: never overlaps - no effect.
Dcm: never overlaps - no effect.
CpG: may overlap - cleavage impaired.
- 5'...m5C GAATTm5C G...3'
|25 to 50% Water, >50% Glycerin, 2.5 to10% Sodium Chloride|
|>90% of DNA fragments can be ligated and re-cut after 50-fold over-digestion with EcoRI|
|Keep container tightly closed|
|10mM Potassium Phosphate (pH 7.4 at 25°C), 300mM NaCl, 1mM EDTA, 1mM DTT, 0.2mg/mL BSA, 0.15% Triton™ X-100, 50% Glycerol|
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