A workflow for the relative quantification of multiple fish species from oceanic water samples using environmental DNA (eDNA) to support large-scale fishery surveys
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Whereas the variety of revealed marine research utilizing environmental DNA (eDNA) has elevated considerably lately, marine fish surveys are nonetheless scarce. To look at the potential for eDNA to help marine fisheries monitoring surveys, we optimized an eDNA isolation methodology, developed a multispecies assay and examined it on eDNA samples collected alongside the Pacific coast of america.
4 industrial DNA extraction kits that exploit the potential of the nucleic acids binding a stable section (two utilizing a silica matrix and two magnetic beads) as nicely an natural separation methodology had been examined. A species-specific multiplex qPCR assay was developed and examined to concurrently goal Pacific hake (Merluccius productus), Pacific lamprey (Entosphenus tridentatus) and eulachon (Thaleichthys pacificus).
The specificity of the assay was examined in silico, in vitro and in natura. Environmental DNA isolation utilizing phenol:chloroform:isoamyl purification with a section lock was optimized and yielded the very best quantity of whole and goal DNA and was used to extract 46 marine water samples for the detection of the three species of curiosity. The multiplex qPCR assay used within the quantification course of was additionally optimized to offer comfort and accuracy.
Pacific hake was current in 44% of the eDNA samples whereas the opposite two species had been absent. Right here, we current an entire workflow for the simultaneous detection and quantification of a number of marine fish species utilizing eDNA. This workflow helps large-scale at-sea sampling efforts with preservation at ambient temperatures and has demonstrated DNA extraction effectivity and reliability. The multiplex qPCR assay is proven to be delicate and particular for the needs of concurrently monitoring the relative abundance of a number of focused fish species.