It is important to use RNase-free water for dissolution of RNA, but not Morpholinos Trace diethylpyrocarbonate (DEPC) left in the water can damage Morpholinos by alkylating bases, but the DEPC can be degraded by autoclaving. Morpholinos are not degraded by RNases, even if the enzyme and oligo are incubated together under favorable conditions for the reaction. Morpholinos have been incubated with rat liver lysates and extracted from animal tissue after in vivo treatment, all without detecting degradation of the Morpholino.
Hudziak RM, Barofsky E, Barofsky DF, Weller DL, Huang SB, Weller DD. Resistance of morpholino phosphorodiamidate oligomers to enzymatic degradation. Antisense Nucleic Acid Drug Dev 1996 Winter;6(4):267-72.
Youngblood DS, Hatlevig SA, Hassinger JN, Iversen PL, Moulton HM. Stability of cell-penetrating Peptide-morpholino oligomer conjugates in human serum and in cells. Bioconjug Chem. 2007 Jan-Feb;18(1):50-60.
Either distilled water or Milli-Q water is likely to be fine for dissolving Morpholinos. It is important that the water be sterile, so autoclave it . We know of instances where the local lab water has some contaminant that causes problems even after distillation or filtration and ion exchange. Testing your water on your cells or in your embryos is a good precaution; if the water is compatible with your biological system, is should be fine with the Morpholino as well.
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