Home FAQs When digesting my dbDNA™ vector with PspOMI (NEB #R0653), the cleavage is incomplete. How can I improve the reaction?

FAQ: When digesting my dbDNA™ vector with PspOMI (NEB #R0653), the cleavage is incomplete. How can I improve the reaction?

PspOMI is a salt-sensitive restriction enzyme. If the sequence of interest (SOI) insert is derived from a low-copy plasmid, a higher input is often required and carryover salts from the plasmid prep can inhibit the restriction enzyme activity. If this is the case, we recommend that the DNA solution added to the reaction be no more than 25% of the total reaction volume to dilute the carryover salts. This can be attained by adjusting the total reaction volume accordingly.