| Publication Type: | Journal Article |
| Year of Publication: | 2020 |
| Authors: | F. Patzold, Zilli, A., Hundsdoerfer, A. K. |
| Journal: | PLoS ONE |
| Volume: | 17 |
| Issue: | 7 |
| Start Page: | e0235222 |
| Pagination: | e0235222 |
| Date Published: | 07/2020 |
| Keywords: | ADNA, COI, COII, DNA, EXTRACTION PROTOCOLS, HYLES, MAXIMUM LIKELIHOOD, METHODOLOGY, MTDNA, MUSEUM, NEXTGEN SEQUENCING, SAMPLING, SPHINGIDAE, TRNA(LEU), TYPES |
| Abstract: | "Here we present and justify an approach for minimal-destructive DNA extraction from historic insect specimens for next generation sequencing applications. An increasing number of studies use insects from museum collections for biodiversity research. However, the availability of specimens for molecular analyses has been limited by the degraded nature of the DNA gained from century-old museum material and the consumptive nature of most DNA extraction procedures. The method described in this manuscript enabled us to successfully extract DNA from specimens as old as 241 years using a minimal-destructive approach. The direct comparison of the DNeasy extraction Kit and the Monarch® PCR & DNA Clean-up Kit showed a significant increase of 17.3-fold higher DNA yield extracted with the Monarch Oligo protocol on average. By using an extraction protocol originally designed for oligonucleotide clean-up, we were able to combine overcoming the restrictions by target fragment size and strand state, with minimising time consumption and labour-intensity. The type specimens used for the minimal-destructive DNA extraction exhibited no significant external change or post-extraction damage, while sufficient DNA was retrieved for analyses." |
| URL: | https://doi.org/10.1371/journal.pone.0235222 |