Thermo Scientific™ NcoI
The NcoI restriction enzyme recognizes C^CATGG sites and cuts best at 37°C in Tango buffer (Isoschizomers: Bsp19I).
Manufacturer: Thermo Scientific™ ER0575
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'...C▵C A T G G...3'
3'...G G T A C▵C...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 37°C
- NcoI 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
Ligation and Recleavage:
- After 50-fold overdigestion with NcoI, more than 95% of the DNA fragments can be ligated and recut
- Dam: never overlaps —no effect
- Dcm: never overlaps —no effect
- CpG: never overlaps —no effect
- EcoKI: never overlaps —no effect
- EcoBI: never overlaps —no effect
Digestion of Agarose-embedded DNA:
- Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours
- AflIII, PscI, BtgI, Eco130I, FatI, PagI
Low salt, high glycerol (>5%) concentrations, pH>8.0 or a large excess of enzyme may result in star activity.