Nhe I

Recognition Sequence:

Heat inactivation: 20 minutes at 65°C.

Ligation and recutting: After 100-fold overdigestion with Nhe I,
>98 % of the DNA fragments can be ligated and recut.

Note: Activity inhibited by salt concentrations > 100mM. Cleaves to leave a 5' CTAG extension which can be efficiently ligated to DNA fragments generated by Avr II, Spe I, or Xba I.

Star activity: Large excess of the enzyme may results in the appearance of star activity.

DNA Methylation:
No Inhibition: dcm, dam
Inhibition (Blocked by some combinations of overlapping): CpG

Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of Lambda DNA (Hind III digest; 1 site) in 1 hour in a total reaction volume of 50 µl. Enzyme activity was determined in the recommended reaction buffer.