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Successful DNA amplification of a more than 200-year-old herbarium specimen: recovering genetic material from the Linnaean era

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Abstract:

The limit for successful DNA extraction was tested by amplification and sequencing of an over 200-year-old herbarium specimen collected by Adam Afzelius, a student of Carl Linnaeus. We amplified and sequenced a 800-bp region between 16S ribosomal DNA and the 3′ part of the trnI gene (16S-trnI) in the chloroplast genome of Phaulopsis talbotii S. Moore (Acanthaceae). To test the replicability and to control for contamination the procedure was performed in sealed vials and with negative PCR controls. The procedure was also repeated in a separate laboratory. In addition, the chloroplast rpl16 intron was successfully amplified and sequenced and the rps16 intron amplified. Sequences of taxa closely related to Acanthaceae were found to be most similar to the produced sequences. The results suggest that molecular investigations of other 18th century botanical collections are feasible and that molecular methods could be employed for comparative studies to extant plant collections. An important application would be to identify descendants or clones of Linnaean lectotypes by comparing DNA from these with potentially remnant plants from Linnaeus' cultivations.

Keywords: 16S-TRNI; CHLOROPLAST DNA; DNA AMPLIFICATION; HERBARIUM SPECIMENS; PHAULOPSIS; RPL16

Document Type: Research Article

Affiliations: 1: Department of Systematic Biology, Evolutionary Biology Centre, Uppsala University, Norbyv. 18D, 75236 Uppsala, Sweden;, Email: katarina.andreasen@ebc.uu.se 2: Department of Systematic Biology, Evolutionary Biology Centre, Uppsala University, Norbyv. 18D, 75236 Uppsala, Sweden 3: Bergius Foundation, Royal Swedish Academy of Sciences, 10405, Stockholm, Sweden, and Department of Botany, Stockholm University, 10 691, Stockholm, Sweden

Publication date: 2009-08-01

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