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Lambda DNA (dam-) digested with MboII, 1.4% agarose, 130 cleavage sitesconventional 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 A G A (N)8▵...3'3'...C T T C T (N)7▵...5'Conditions for 100% Activity:1X Buffer B: 10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2 and 0.1mg/mL BSAIncubate at 37°CStorage Buffer:MboII 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) glycerolMethylation Effects:Dam: may overlap —blockedDcm: never overlaps —no effectCpG: may overlap —no effectEcoKI: never overlaps —no effectEcoBI: may overlap —no effect Note:Assayed using lambda DNA (dam) (#SD0021). MboII is blocked by overlappingdam methylation. To avoiddam methylation, use adam,dcm- strain such as GM2163 (#M0099). Greater than 15-fold overdigestion with MboII may result in star activity. MboII produces DNA fragments that have a single-base 3 extension which are more difficult to ligate than blunt-ended fragments. MboII 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.
