Thermo Scientific™ PaeI (SphI)
The PaeI (SphI) restriction enzyme recognizes GCATG^C sites and cuts best at 37°C in B buffer (Isoschizomers: BbuI, SphI).
Manufacturer: Thermo Scientific™ ER0602
Lambda DNA digested with PaeI (SphI), 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 A T G▵C...3'
3'...C▵G T A C G...5'
Conditions for 100% Activity:
- 1X Buffer B: 10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2 and 0.1mg/mL BSA
- Incubate at 37°C
- PaeI is supplied in: 10mM potassium-phosphate (pH 7.0 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.15% Triton X-100, 0.5mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with PaeI, more than 95% of the DNA fragments can be ligated and recut
- Dam: never overlaps —no effect
- Dcm: never overlaps —no effect
- CpG: may overlap —no effect
- EcoKI: never overlaps —no effect
- EcoBI: may overlap —blocked
Digestion of Agarose-embedded DNA:
- Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours
- Hin1II, XceI