The use of herbarium specimens in DNA phylogenetics: Evaluation and improvement

Article Article scientifique Article The use of herbarium specimens in DNA phylogenetics: Evaluation and improvement eng 1995 author Savolainen Vincent author Cuenoud Philippe 26865 author Spichiger Rodolphe author Martinez Maria D. P. author Crèvecoeur Michèle 22928 author Manen Jean-François 87-98 10.1007/BF00984634 0378-2697 During the last few years we have been confronted with the need to use herbarium specimens in the molecular phylogeny studies, since it is generally difficult to obtain living material of some rare species. Ancient DNA has been sequenced, and there are also reports on successful DNA amplification from herbarium specimens. However, it is not easy to obtain amplified DNA from the first herbarium sample tested. In this paper, experiments are described about trials of DNA amplification from two to 151-year-old herbarium specimens of plant species we needed for our projects. Of the 17 herbarium samples tested only two allowed DNA amplification under standard DNA isolation conditions. Different types of PCR inhibiting activities were demonstrated in DNA extracts. In some of the extracts there was extremely low concentration of template with satisfactory quality. In some instances, PCR inhibiting activities were successfully removed by treating them either with insoluble polyvinylpyrrolidone or by adding bovine serum albumin (BSA) to the amplification mixture. However, some PCR-inhibiting activities were resistant to the treatments described above. When the concentration of template was very low, a second PCR amplification with internal primers was necessary to increase the amount of DNA for sequencing. Nevertheless, contamination of either DNA extract or amplification mixture were sometimes observed, and consequently precautions were taken to minimize them. Finally, successful amplification was obtained in eight samples out of the 17 examined. Herbarium specimens Molecular phylogenetics Ancient DNA PCR techniques Dewey 580 45 Département des sciences végétales 163 false Plant systematics and evolution 197 1-4 42b6dcd8-51b1-4772-912e-ed6c4247f6b4 Article (Published version) Public Savolainen1995_Article_TheUseOfHerbariumSpecimensInDN.pdf 2000268 application/pdf openAccess Faculté des sciences / Section de biologie / Département des sciences végétales

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