Thermo Scientific™ FspAI
The FspAI restriction enzyme recognizes RTGC^GCAY sites and cuts best at 37°C in O buffer.
Manufacturer: Thermo Scientific™ ER1661
Lambda DNA digested with FspAI, 0.5% agarose, 2 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.
More than 80% of the digested DNA fragments can be ligated and more than 90% can be recut at a 5ft.-termini concentration of 0.2μM after a 50-fold overdigestion (3U/μg DNA x 17 hours) with FspAI.
5'...R T G C▵G C A Y...3'
3'...Y A C G▵C G T R...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
- FspAI 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
- Dam: never overlaps — no effect
- Dcm: never overlaps — no effect
- CpG: completely overlaps — blocked
- EcoKI: may overlap — effect not determined
- EcoBI: may overlap — effect not determined
Low salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity.