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
Inactivation Temperature (20 min): 80°C
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.
– 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.
All preparations are assayed for contaminating endonuclease, 3′-exonuclease, and nonspecific single- and double-stranded DNase activities.