Thermo Scientific™ Eco57I (AcuI)
The Eco57I (AcuI) restriction enzyme recognizes CTGAAG(16/14)^ sites and cuts best at 37°C in G (+SAM) buffer (Isoschizomers: AcuI).
Marca: Thermo Scientific™ ER0341
Detalles adicionales : Peso : 1.51950kg
5'...C T G A A G (N)16▵...3'
3'...G A C T T C (N)14▵...5'
Conditions for 100% Activity:
- [1X Buffer G] + SAM:[10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, 50mM NaCl and 0.1mg/mL BSA] + 0.01mM S-adenosylmethionine
- Incubate at 37°C
- Eco57I is supplied in:10mM potassium phosphate (pH 7.4 at 25°C), 100mM NaCl, 1mM EDTA, 1mM DTT and 50% (v/v) glycerol
Ligation and Recleavage:
- After 10-fold overdigestion with Eco57I, approximately 70% of the DNA fragments can be ligated, but none of these can be recut due to the methylation of the recognition sequence by this enzyme
- Dam: never overlaps-no effect
- Dcm: never overlaps-no effect
- CpG: never overlaps-no effect
- EcoKI: never overlaps-no effect
- EcoBI: may overlap-effect not determined
Eco57I requires only Mg2 + for its activity, but is stimulated by S-adenosylmethionine. 0.01mM S-adenosylmethionine gives a 100-fold increase in Eco57I activity. Still, complete cleavage of some substrates with Eco57I is difficult to achieve. Eco57I concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase. Low salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity. Eco57I may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.