000K utf8 1100 2021$c2021-06-11 1500 eng 2050 urn:nbn:de:hbz:465-20240814-132338-8 2051 10.1002/ece3.7753 3000 Buchner, Dominik 3010 Beermann, Arne J. 3010 Leese, Florian 3010 Weiss, Martina 4000 Cooking small and large portions of “biodiversity-soup”$dMiniaturized DNA metabarcoding PCRs perform as good as large-volume PCRs [Buchner, Dominik] 4209 DNA metabarcoding is a powerful tool to assess arthropod diversity in environmental bulk samples such as Malaise trap, pitfall trap, or hand net samples. While comparative performance tests for different extraction protocols, primers, and Taq polymerases have been made, the effect of different PCR volumes on bulk sample metabarcoding performance is less explored. Although using small PCR volumes reduces overall costs, they may lead to decreased taxon recovery or higher replicate variability due to increased pipetting imprecision, PCR stochasticity (PCR drift), or inhibition when using high amounts of template community DNA. We here performed a simple DNA metabarcoding experiment to test if species detection and the consistency of technical replicates decrease with decreasing PCR volume in standard reaction tubes. We used a mock community sample consisting of different amounts of DNA from 35 arthropod species, and a Malaise trap sample composed of many thousand insect specimens. PCR volumes tested were 5, 10, 15, 20, 25, and 50 µl. Both samples were replicated 14 times in the first PCR step with two technical replicates each in the second PCR step. Our data show that small PCR volumes did neither have systematically lower species detection or richness values, nor lower consistency between PCR replicates. We therefore recommend low volumes primarily depending on handling constraints. Further, we emphasize the importance of sequencing depth for taxon recovery. 4950 https://doi.org/10.1002/ece3.7753$xR$3Volltext$534 4950 https://nbn-resolving.org/urn:nbn:de:hbz:465-20240814-132338-8$xR$3Volltext$534 4961 https://duepublico2.uni-due.de/receive/duepublico_mods_00081889 5051 570 5550 amplicon sequencing 5550 biomonitoring 5550 bulk sample 5550 Malaise trap 5550 PCR bias