Inter-laboratory ring test for environmental DNA extraction protocols: implications for marine megafauna detection using three novel qPCR assays DOI Creative Commons
Lauren Rodriguez, Lorenzo De Bonis, J.S. McKee

et al.

Metabarcoding and Metagenomics, Journal Year: 2025, Volume and Issue: 9

Published: Jan. 7, 2025

The comparability of methods applied to environmental DNA (eDNA) samples across laboratories remains a significant challenge for biodiversity monitoring on global scale. Performance differences between protocols can jeopardize effective conservation strategies regions and focal species. To address potential discrepancies amongst four international partners within collaborative eDNA initiative, an inter-laboratory comparison (i.e., ring test) was conducted compare efficiencies established extraction methodologies based 39 samples. Each laboratory contributed eight eleven collected throughout the North-East Atlantic Mediterranean Sea near sperm whales, porbeagle sharks, basking bottlenose dolphins common dolphins. After lysis, aliquots were exchanged before subsequent using each facility’s preferred method. Extracts returned lysates’ respective origin measurements total concentration, as well quantitative PCR three novel species-specific assays marine megafauna. Our findings revealed similar concentrations DNA, yet reduction in performance targeted qPCR reactions by one laboratory, who has therefore modified their method be used remainder this project. Overall, detection success differed target taxa with sharks being less often detected (and at lower concentrations) than mammals. Significant interaction effects found combinations species, suggesting link variable conditions. study serves foundational step towards establishing reproducible practices that are crucial multinational projects enable comparable results.

Language: Английский

Inter-laboratory ring test for environmental DNA extraction protocols: implications for marine megafauna detection using three novel qPCR assays DOI Creative Commons
Lauren Rodriguez, Lorenzo De Bonis, J.S. McKee

et al.

Metabarcoding and Metagenomics, Journal Year: 2025, Volume and Issue: 9

Published: Jan. 7, 2025

The comparability of methods applied to environmental DNA (eDNA) samples across laboratories remains a significant challenge for biodiversity monitoring on global scale. Performance differences between protocols can jeopardize effective conservation strategies regions and focal species. To address potential discrepancies amongst four international partners within collaborative eDNA initiative, an inter-laboratory comparison (i.e., ring test) was conducted compare efficiencies established extraction methodologies based 39 samples. Each laboratory contributed eight eleven collected throughout the North-East Atlantic Mediterranean Sea near sperm whales, porbeagle sharks, basking bottlenose dolphins common dolphins. After lysis, aliquots were exchanged before subsequent using each facility’s preferred method. Extracts returned lysates’ respective origin measurements total concentration, as well quantitative PCR three novel species-specific assays marine megafauna. Our findings revealed similar concentrations DNA, yet reduction in performance targeted qPCR reactions by one laboratory, who has therefore modified their method be used remainder this project. Overall, detection success differed target taxa with sharks being less often detected (and at lower concentrations) than mammals. Significant interaction effects found combinations species, suggesting link variable conditions. study serves foundational step towards establishing reproducible practices that are crucial multinational projects enable comparable results.

Language: Английский

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