Abstract
At the same time that molecular researchers are improving techniques to extract DNA from museum specimens, this increased demand for access to museum specimens has created tension between the need to preserve specimens for maintaining collections and morphological research and the desire to conduct molecular analyses. To address these concerns, we examined the suitability of non-invasive DNA extraction techniques on three species of parasitic Hymenoptera (Braconidae), and test the effects of body size (parasitoid species), age (time since collection), and DNA concentration from each extract on the probability of amplifying meaningful fragments of two commonly used genetic loci. We found that age was a significant factor for determining the probability of success for sequencing both 28S and COI fragments. While the size of the braconid parasitoids significantly affected the total amount of extracted DNA, neither size nor DNA concentration were significant factors for the amplification of either gene region. We also tested several primer combinations of various lengths, but were unable to amplify fragments longer than ∼150 base pairs. These short fragments of 28S and COI were however sufficient for species identification, and for the discovery of within species genetic variation.
Highlights
Methods for extracting and analyzing DNA sequence data from specimens not immediately preserved for DNA extraction are improving at a rapid rate, as highlighted by the recent sequencing of the Neanderthal genome [1]
Longifemoralis, 9 specimens in the genus Meteorus, and 12 specimens of T. pallidus, with specimens ranging in age at time of extraction from 1 to 96 years
Results from the ANCOVA analysis showed that the total amount of genomic material (DNA concentration) differed significantly between parasitoid species
Summary
Methods for extracting and analyzing DNA sequence data from specimens not immediately preserved for DNA extraction are improving at a rapid rate, as highlighted by the recent sequencing of the Neanderthal genome [1]. Among these methods, several techniques exist which allow DNA to be extracted from a specimen without conferring visible damage [2,3,4]. DNA extracted from museum specimens has been a useful source of information for understanding recent shifts in population structure, especially with regard to population declines in native pollinators [10,11], in addition to having been helpful in the context of molecular based identifications [3], and the short fragments of DNA extracted from museum specimens have recently been used in Next-Generation Sequencing applications [12]
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