Thermo Scientific™ PagI (BspHI)
The PagI (BspHI) restriction enzyme recognizes T^CATGA sites and cuts best at 37°C in O buffer (Isoschizomers: BspHI, CciI, RcaI).
Marca: Thermo Scientific™ ER1281
Detalles adicionales : Peso : 0.00900kg
5'...T▵C A T G A...3'
3'...A G T A C▵T...5'
Conditions for 100% Activity:
- 1X Buffer O: 50mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, 100mM NaCl and 0.1mg/mL BSA
- Incubate at 37°C
- PagI is supplied in: 10mM Tris-HCl (pH 7.4 at 25°C), 100mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with PagI, more than 95% of the DNA fragments can be ligated and recut
- Dam: may overlap —cleavage impaired
- Dcm: never overlaps —no effect
- CpG: never overlaps —no effect
- EcoKI: never overlaps —no effect
- EcoBI: may overlap —cleavage impaired
Digestion of Agarose-embedded DNA:
- Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours
- AflIII, PscI, DsaI, Eco130I, NcoI
Low salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity. PagI cleavage is impaired by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm strain such as GM2163 (#M0099).