Archived otoliths constitute an important source of historical DNA for use in temporal genetic studies, but such otoliths are also valuable for other research applications, e.g. growth or microchemistry studies, where information about the past is of relevance. Consequently, there are potentially conflicting interests regarding how the limited and irreplaceable otolith collections should be used. To resolve this, it is important to find out whether DNA extraction damages otoliths such that they can no longer be used for other research purposes or whether individual otoliths can be used in multiple applications. We examined the effects of three different DNA extraction methods on the elemental composition, the morphology, and the clarity of annual growth increments for successful age estimation of Atlantic cod (Gadus morhua) otoliths that had been archived for 0–31 years. The three extraction methods yielded DNA of comparable quality, and none of the methods caused major damage to the otoliths. Of the element concentrations measured, only Mg and Rb showed considerable changes resulting from DNA extraction. The physical properties of the otolith (morphology and clarity of annual growth increments) were not affected. Hence, cod otoliths can be used for several research purposes after DNA extraction.