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

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)