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


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1995-03

AUTHORS

Vincent Savolainen, Philippe Cuénoud, Rodolphe Spichiger, Maria D. P. Martinez, Michèle Crèvecoeur, Jean-François Manen

ABSTRACT

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. More... »

PAGES

87-98

Journal

TITLE

Plant Systematics and Evolution

ISSUE

1-4

VOLUME

197

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00984634

DOI

http://dx.doi.org/10.1007/bf00984634

DIMENSIONS

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