Eco57I (AcuI)

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

• Eco57I requires only Mg²⁺ for its activity, but is stimulated by S-adenosylmethionine. 0.01 mM 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.

For methylation sensitivity refer to product specifications.

Reaction conditions

Methylation Effects

• 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.

Cleavage efficiency close to the termini of PCR fragments

Number of recognition sites in DNA molecules