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This document specifies procedures for extracting environmental DNA (eDNA) that has been captured on filters after filtering water from aquatic environments. This document applies to the eDNA from any target taxonomic group that has been captured on filters, from single species to whole communities. It applies to eDNA in both intracellular (i.e., within cells, either with or without a cell wall) and extracellular (i.e., free or attached to particles of biological or non-biological origin) form. Procedures in this document do not apply to the extraction of eDNA from other starting materials, including but not limited to biofilms, sediments, and bulk samples.
This standard is designed for eDNA extractions performed in a laboratory and is not designed for fieldbased eDNA extractions. This document does not include procedures for water sampling, filtering, and sample preservation and storage after filtering; these are covered by ISO FDIS 17805. This document provides the following procedures and/or guidelines as they relate to eDNA extraction from filters: (1) contamination prevention; (2) eDNA extraction method selection; (3) positive and negative controls; (4) lysis and purification steps; (5) quantifying and storing eDNA extracts; and (6) quality testing of extracted eDNA for downstream applications, with a focus on PCR-based biodiversity monitoring.
Aquatic macroinvertebrates are the key organismal group required for regulatory biomonitoring of water bodies world-wide (e.g. European Water Framework Directive, 2000/60/EG, South African National Water Act, Australian River Assessment System, National Censuses on River Environments (NCRE) in Japan, US Clean Water Act) and environmental impact assessment. They are also a key group for ecotoxicological assessments as well as in the context of global biodiversity monitoring (e.g. EU biodiversity strategy, Kunming-Montreal Global Biodiversity Framework). With DNA-based approaches, such as DNA metabarcoding, the taxonomic composition of whole aquatic macroinvertebrate communities can be assessed, yet for comparable results standardisation of procedures is essential, including, but not limited to, maximisation of DNA quality, avoidance of contamination of DNA from other sources.
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