DpnI

Source: Diplococcus pneumoniae

5′-G A T C- 3′
3′-C T A G- 5′

Purified from E.coli strain that carries the DpnI gene from Diplococus pneumoniae.

Reaction Temperature: 37°C

Prototype: DpnI

Inactivation Temperature (20 min): 80°C

Reaction Buffer:

1x ONE Buffer

Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of pBR322 dam methylated DNA in 1 hr. Total reaction volume is 50 µl.

Notes: 
– DpnI cleaves only methylated GATC sites;
– it may be necessary to add more enzyme to obtain complete digestion when using other buffer than optimal (ONE Buffer).
– it is recommended to add 10 u of the enzyme to obtain complete digestion of the methylated template after standard site-directed mutagenesis (total reaction volume is 50 µl)

Storage Conditions: Store at –20°C.

Quality Control:

All preparations are assayed for contaminating endonuclease, 3′-exonuclease, and nonspecific single- and double-stranded DNase activities.