Thermo Scientific™ BstXI
Cut at CCANNNNN^NTGG sites with BstXI restriction enzyme, which performs best at 55°C in O buffer.
Manufacturer: Thermo Scientific™ ER1022
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 C A N N N N N▵N T G G...3'
3'...G G T N▵N N N N N A C C...5'
Conditions for 100% Activity:
- b1X Buffer O: 50mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, 100mM NaCl and 0.1mg/mL BSA
- Incubate at 55°C
- BstXI 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 BstXI, approximately 95% of the DNA fragments can be ligated and recut
- Dam: never overlaps — no effect
- Dcm: may overlap — cleavage impaired
- CpG: never overlaps — no effect
- EcoKI: never overlaps — no effect
- EcoBI: never overlaps — no effect
Digestion of Agarose-embedded DNA:
- Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours
Incubation at 37°C results in 50% activity. Greater than 15-fold overdigestion with BstXI may result in star activity. BstXI cleavage is impaired by overlappingdcm methylation. To avoiddcm methylation, use adam-, dcm strain such as GM2163 (#M0099).