Cut at GGATC(4/5)^ sites with BspPI (AlwI) restriction enzyme, which performs best at 55°C in Tango buffer (Isoschizomers: AclWI, AlwI).
Brand: Thermo Scientific ER1322
Code : 88
Additional Details : Weight : 0.00900kg
5'...G G A T C (N)4▵...3'
3'...C C T A G (N)5▵...5'
Conditions for 100% Activity:
- 1X Buffer Tango™: 33mM Tris-acetate (pH7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA
- Incubate at 55°C
- BspPI is supplied in:10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 10-fold overdigestion with BspPI, approximately 70% of the DNA fragments can be ligated in a reaction mixture containing 20-40U of T4 DNA Ligase/1μg of fragments and 10% PEG. More than 50% of these can be recut
- Dam: completely overlaps — blocked
- Dcm: may overlap — no effect
- CpG: may overlap — no effect
- EcoKI: never overlaps — no effect
- EcoBI: may overlap — effect not determined
Incubation at 37°C results in 30% activity. Assayed using lambda DNA (dam) (#SD0021). BspPI is blocked by overlappingdam methylation. To avoiddam methylation, use adam-, dcm strain such as GM2163 (#M0099).
|1 to 3U/µL|