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Ukr. Bot. J. 2018, 75(6): 564–575
https://doi.org/10.15407/ukrbotj75.06.564
Plant Physiology, Biochemistry, Cell and Molecular Biology

DNA extraction from old herbarium material of Veronica subgen. Pseudolysimachium (Plantaginaceae)

Höpke J.1, Brewer G.2, Dodsworth S.2,3, Ortiz E.M.4, Albach D.C.1
Abstract

Herbarium specimens have become a major source of information in molecular biodiversity research, framing the term "herbarium genomics". However, obtaining good DNA from old herbarium specimens is still a challenge. Currently, DNA extraction methods from old herbarium material often yield highly degraded and fragmented DNA. A number of studies have discussed such methods, especially how to avoid further DNA fragmentation. This study aims to compare different DNA extraction methods applied to old herbarium material from Veronica subg. Pseudolysimachium. One such method is a CTABbased DNA extraction followed by a clean-up with paramagnetic beads that is used in the Jodrell Laboratory, Royal Botanic Gardens Kew, UK. This method was compared to a modified NucleoSpin Plant II protocol, based on silica columns, as used at the Technical University Munich-Freising, which was already successfully used for extracting DNA from a Linnean type specimen. Further tests were conducted on the influence of incubation time on the CTAB DNA extraction protocol with a subsample of specimens. Our preliminary results suggest that CTAB DNA extraction might have some advantages in specific cases but also that silica column-based methods have fewer problems with contamination by polysaccharides and polyphenolic compounds. Regarding the incubation time, we did not observe a clear pattern, but we developed several ideas on how to proceed with tests to find an optimal DNA extraction protocol to deal with highly fragmented DNA. Taking practical considerations into account, the column-based method proves to be preferable, especially when trying to reduce the amount of leaf tissue used, but further modifications of both methods should be explored.

Supplementary Material. Electronic Supplement (Table E1, p. e3) is available in the online version of this article below.

Keywords: Veronica subg. Pseudolysimachium, herbarium specimens, DNA extraction methods, molecular biodiversity research

Full text: PDF (Eng) 2.09M

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