Thermo Scientific™ KpnI
The KpnI restriction enzyme recognizes GGTAC^C sites and cuts best at 37°C in its own unique buffer (Isoschizomers: Asp718I).
Manufacturer: Thermo Scientific™ ER0523
Lambda DNA digested with KpnI, 0.7% 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.
5'..G G T A C▵C...3'
3'..C▵C A T G G...5'
Conditions for 100% Activity:
- 1X Buffer KpnI: 10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, 0.02% Triton X-100 and 0.1mg/mL BSA
- Incubate at 37°C
- KpnI is supplied in: 10mM Tris-HCl (pH 7.5 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
- Dam: never overlaps —no effect
- Dcm: may overlap —no effect
- CpG: may overlap —no effect
- 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
High glycerol (>5%) concentrations, pH>8.0 or a large excess of enzyme may result in star activity.