The NcoI restriction enzyme recognizes C^CATGG sites and cuts best at 37°C in Tango buffer (Isoschizomers: Bsp19I).
Brand: Thermo Scientific ER0571
Code : 88
Additional Details : Weight : 0.00900kg
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.