Thermo Scientific™ NotI
The NotI restriction enzyme recognizes GC^GGCCGC sites and cuts best at 37°C in O buffer (Isoschizomers: CciNI).
Marca: Thermo Scientific™ ER0592
Detalles adicionales : Peso : 1.51950kg
5'..G C▵G G C C G C...3'
3'..C G C C G G▵C G...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
- NotI is supplied in: 20mM Tris-HCl (pH 7.8 at 25°C), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 0.02% Triton X-100, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with NotI, more than 95% of DNA fragments can be ligated and recut
- Dam: never overlaps —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 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded Adenovirus-2 DNA in 16 hours
- Bsp120I, CfrI, Eco52I
Assayed using pTZ19RJL2 plasmid DNA digested with BseLI. Supercoiled plasmids may require up to 5-fold more NotI for complete digestion than linear DNAs.