Thermo Scientific™ Kpn2I (BspEI)
The Kpn2I (BspEI) restriction enzyme recognizes T^CCGGA sites and cuts best at 55°C in Tango buffer (Isoschizomers: AccIII, Aor13HI, BseAI, Bsp13I, BspEI, MroI).
Manufacturer: Thermo Scientific™ ER0531
Lambda DNA digested with Kpn2I (BspEI), 0.7% agarose, 24 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'..T▵C C G G A...3'
3'..A G G C C▵T...5'
Conditions for 100% Activity:
- 1X Buffer Tango™: 33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA
- Incubate at 55°C
- Kpn2I 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: may overlap —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 5U of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours
- BshTI, BsaWI, Cfr9I, Cfr10I, Eco88I, NgoMIV, SgrAI
Incubation at 37°C results in 50% activity.