Thermo Scientific™ NotI (10 U/µL) Thermo Scientific™ ER0593

Description

The NotI restriction enzyme recognizes GC^GGCCGC sites and cuts best at 37°C in O buffer (Isoschizomers: CciNI).

Description

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

Storage Buffer:

• 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

Methylation Effects:

• 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

Compatible Ends:

• Bsp120I, CfrI, Eco52I

Note:

Assayed using pTZ19RJL2 plasmid DNA digested with BseLI. Supercoiled plasmids may require up to 5-fold more NotI for complete digestion than linear DNAs.