Thermo Scientific™ GsuI (BpmI) (5 U/μL)

Description

Lambda DNA digested with GsuI (BpmI), 1% agarose, 25 cleavage sites

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.

5'...C T G G A G (N)₁₆▵...3'

3'...G A C C T C (N)₁₄▵...5'

Conditions for 100% Activity:

• 1X Buffer B: 10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl₂ and 0.1mg/mL BSA
• Incubate at 30°C

Storage Buffer:

• GsuI is supplied in:10mM potassium phosphate (pH 7.5 at 25°C), 1mM EDTA, 1mM DTT, 0.2mg/mL BSA and 50% (v/v) glycerol

Methylation Effects:

• Dam: never overlaps —no effect
• Dcm: may overlap —blocked
• CpG: never overlaps —no effect
• EcoKI: never overlaps —no effect
• EcoBI: never overlaps —no effect

Note:

• Incubation at 37°C results in 70% activity
• GsuI requires only Mg² + for its activity, but is stimulated by S-adenosylmethionine
• 0.01mM S-adenosylmethionine gives a 2-fold increase in activity
• GsuI 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 and SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis
• GsuI is blocked by overlappingdcm methylation
• To avoiddcm methylation, use adam,dcm- strain such as GM2163 (#M0099)