Thermo Scientific™ MboI
The MboI restriction enzyme recognizes ^GATC sites and cuts best at 37°C in R buffer (Isoschizomers: BfuCI, BssMI, BstKTI, BstMBI, DpnII, Kzo9I, NdeII, Sau3AIm).
Manufacturer: Thermo Scientific™ ER0812
Lambda DNA (dam-) digested with MboI, 1.4% agarose, 116 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'..▵G A T C ...3'
3'.. C T A G▵...5'
Conditions for 100% Activity:
- 1X Buffer R: 10mM Tris-HCl (pH 8.5 at 37°C), 10mM MgCl2, 100mM KCl and 0.1mg/mL BSA
- Incubate at 37°C
- MboI is supplied in:10mM Tris-HCl (pH 7.5 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with MboI, more than 95% of the DNA fragments can be ligated and recut
- Dam: completely overlaps —blocked
- Dcm: never overlaps —no effect
- CpG: may overlap —no effect
- EcoKI: never overlaps —no effect
- EcoBI: may overlap —blocked
Assayed using lambda DNA (dam) (#SD0021). MboI is blocked by overlappingdam methylation. To avoiddam methylation, use adam,dcm- strain such as GM2163 (#M0099). MboI, Bsp143I and DpnI all recognize the same sequence but have different methylation sensitivities and cleavage sites.