Not I

Recognition Sequence:

Heat inactivation: 20 minutes at 65°C.

Ligation and recutting: After 25-fold overdigestion with Not I,
>98% of the DNA fragments can be ligated and recut with this enzyme.

Note: Supercoiled plasmids may require up to 5-fold more Not I for complete digestion than linear DNA.

DNA Methylation:
No Inhibition: dam, dcm
Inhibition (Blocked): CpG

Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of Adenovirus-2 DNA (7 sites) in 1 hour in a total reaction volume of 50 µl. Enzyme activity was determined in the recommended reaction buffer.