Thermo Scientific™ BstXI
Cut at CCANNNNN^NTGG sites with BstXI restriction enzyme, which performs best at 55°C in O buffer.
Brand: Thermo Scientific™ ER1021
Code : 88
Additional Details : Weight : 1.51950kg
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).