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Storage of Morpholinos: refrigerate or room temperature?

Morpholino oligos are completely chemically & biologically (enzymatically) stable sitting in water at 4C or at room temperature as long as the pH is not extreme. However, they undergo physical changes that can affect their biological activity. Chilled storage increases the tendency of the oligos to associate with container walls (this is especially a problem if the oligos undergo freeze-thaw cycles). Once a Morpholino has associated tightly with the wall of a container, it can be difficult or impossible to get the oligo back into solution at neutral pH. Very acidic conditions can protonate the oligo and get it back into the bulk solution phase, but can also damage the Morpholino’s backbone, usually by hydrolyzing off the dimethylamine; this is useful for assessing the presence of Morpholinos on the walls of a dry tube, but not for recovering biological activity of a stock solution that has underwent wall-association. Because of this, we do not recommend storing Morpholinos at 4C.

At room temperature, wall association occurs less but solution-phase aggregation occurs more frequently. This aggregation can decrease the single-stranded antisense activity of a Morpholino solution. However, the biological activity of the solution can usually be recovered by autoclaving. Faced with a choice between the irrecoverable loss of activity though container wall association at low temperature versus the recoverable loss of activity through aggregation at room temperature, we recommend room temperature storage.

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