Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens

PLoS One. 2020 Jul 8;15(7):e0235222. doi: 10.1371/journal.pone.0235222. eCollection 2020.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA / genetics
  • DNA / isolation & purification*
  • Fossils / anatomy & histology
  • High-Throughput Nucleotide Sequencing
  • Insecta / anatomy & histology
  • Insecta / chemistry
  • Insecta / genetics*
  • Phylogeny

Substances

  • DNA

Grants and funding

This work of FP and AH was financed by the Deutsche Forschungsgemeinschaft (DFG HU 1561/5-1 with support from RE 603/25-1) and benefitted from the sharing of expertise within the DFG priority program SPP 1991 Taxon-Omics. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. https://www.dfg.de/gefoerderte_projekte/programme_und_projekte/listen/projektdetails/index.jsp?id=313688472