Repeated subsamples during DNA extraction reveal increased diversity estimates in DNA metabarcoding of Malaise traps

• DOI: 10.1002/ece3.9502
• Publication Date: 2022-11-28T06:52:35Z
• Authors: Vera M. A. Zizka; Matthias F. Geiger; Thomas Hörren; Ameli Kirse; Niklas W. Noll; Livia Schäffler; Alice M. Scherges; Martin Sorg
• Keywords: Homogenization

With increased application of DNA metabarcoding in biodiversity assessment, various laboratory protocols have been optimized, and their further evaluation is subject current research. Homogenization bulk samples subsequent extraction from a subsample destructed tissue common first stage the process. This can either be conducted using sample material soaked storage fixative, e.g., ethanol (here referred to as "wet" treatment) or dried individuals ("dry"). However, it remains uncertain if perfect mixing equal distribution within tube ensured during homogenization what extent incomplete resulting variations composition affect diversity assessments only fraction processed downstream workflow. Here we investigated efficiency under wet dry conditions tested how might complex arthropod samples. We considered five time intervals Malaise trap process nine different subsamples homogenized (20 mg each) both treatments. Results indicate more consistent assessment material, but at cost higher processing time. Both approaches detected comparable OTU revealed similar taxa compositions single extraction. an number extraction, for approaches, especially highly diverse obtained summer. Here, particularly detection small low-biomass increased. The multiple protocol therefore helpful procedure enhance estimates counteract taxonomic bias assessments. induces costs effort large-scale monitoring schemes needs on project-specific prospects.