Coupling non‐destructive DNA extraction and voucher retrieval for small soft‐bodied Arthropods in a high‐throughput context: the example of Collembola

Abstract

Here, we describe a simple method adapted for high-throughput protocols allowing voucher specimen recovery for Collembola and by extension for other soft-bodied small arthropods. A standard extraction protocol was tested to examine the effects of lysis duration (1, 2, 4, 12 h) on DNA concentration, amplification success and specimen condition. Good quality DNA was obtained after 1 h of lysis, while voucher condition was fine for up to 12 h. The lysis step substantially shortens the clearing process necessary for morphological examination.

Biological invasions in soil: DNA barcoding as a monitoring tool in a multiple taxa survey targeting European earthworms and springtails in North America

“…Such DNA barcode analyses will enable large-scale studies requiring substantial numbers of routine identifications. The ability of DNA barcodes to distinguish closely related species in earthworms and Collembola has been established in several prior studies (Hogg and Hebert 2004;Chang et al 2009;Richard et al 2010;Porco et al 2010b). However, this approach also brings a finer resolution than morphology by revealing potential cases of cryptic diversity in well studied species (Table 1).…”

DNA Barcodes for Marine Biodiversity: Moving Fast Forward?

“…Recent developments provide non-invasive DNA extraction with total voucher recovery [97], as well as extraction of DNA leaked into the aquatic environment [98] or ethanol [33]. Primers are being developed for various taxa and additional markers or larger COI fragments used in cases of slow mutation rate (e.g., sponges, cnidarians).…”

Four new species, DNA barcode library and pre-Pliocene speciation of the euedaphic Afromontane Clivinini genera Trilophidius and Antireicheia (Coleoptera: Carabidae, Scaritinae)

“…Specimens were preserved in 96% ethanol and processed for downstream DNA extraction and sequencing (Hebert et al 2003;Ratnasingham & Hebert 2007). Genomic DNA was extracted from either a single leg, or (considering the small size of specimens) following the whole-body non-destructive protocol developed for Collembola (Porco et al 2010). DNA extraction, purification, amplification and sequencing was performed in a commercial laboratory "Canadian Center for DNA Barcode" (CCDB, http://www.ccdb.ca/) at the University of Guelph, Ontario, Canada following standard protocol (Ivanova et al 2006).…”