Thermo Scientific™ PauI (BssHII)
The PauI (BssHII) restriction enzyme recognizes G^CGCGC sites and cuts best at 37°C in R buffer (Isoschizomers: BsePI, BssHII).
Manufacturer: Thermo Scientific™ ER1091
Lambda DNA digested with PauI (BssHII), 0.7% agarose, 6 cleavage sites
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 G C G C...3'
3'...C G C G C▵G...5'
Conditions for 100% Activity:
- 1X Buffer R: 10mM Tris-HCl (pH 8.5 at 37°C), 10mM MgCl2, 100mM KCl and 0.1mg/mL BSA
- Incubate at 37°C
- PauI is supplied in:10mM Tris-HCl (pH 7.5 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mLBSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with PauI, more than 95% of the DNA fragments can be ligated and recut
- Dam: never overlaps —no effect
- Dcm: never overlaps —no effect
- CpG: completely overlaps —blocked
- EcoKI: never overlaps —no effect
- EcoBI: never overlaps —no effect
Digestion of Agarose-embedded DNA:
- Minimum 10units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours
- AflIII, AscI, BtgI, MluI
Low salt, high glycerol (>5%) concentrations or a large excess of enzyme may result in star activity.