Thermo Scientific BseXI (BbvI)
Cut at GCAGC(8/12)^ sites with BseXI (BbvI) restriction enzyme, which performs best at 65°C in its own unique buffer (Isoschizomers: BbvI, BstV1I).
Manufacturer: Thermo Scientific ER1451
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 C A G C (N)8▵...3'
3'...C G T C G (N)12▵...5'
Conditions for 100% Activity:
- 1X Buffer BseXI: 50mM Tris-HCl (pH 7.5 at 37°C), 2mM MgCl2, 100mM NaCl and 0.1mg/mL BSA
- Incubate at 65°C
- To ensure higher efficiency of digestion, perform the cleavage reaction under paraffin oil
- BseXI is supplied in:10mM Tris-HCl (pH 7.4 at 25°C), 200mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 10-fold overdigestion with BseXI, more than 95% of the DNA fragments can be ligated recut
- Dam: never overlaps no effect
- Dcm: never overlaps no effect
- CpG: may overlap no effect
- EcoKI: never overlaps no effect
- EcoBI: may overlap effect not determined
Assayed using pBR322 DNA (#SD0041). Incubation at 37°C results in 10% activity. Greater than 40-fold overdigestion with BseXI may result in star activity. BseXI 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.
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