Thermo Scientific™ NotI (10 U/μL)

Description

Ad2 DNA digested with NotI, 0.7% agarose, 7 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 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.