Capture of target template with biotinylated probes from eDNA samples

Environmental DNA (eDNA) metabarcoding from water samples has shown great promise for biodiversity monitoring in recent years. However, universal primers targeting the cytochrome oxidase I (COI) marker gene popular in metazoan studies have displayed high levels of nontarget amplification. To date, enrichment methods bypassing amplification have not been able to match the detection levels of conventional metabarcoding. This study evaluated the use of universal metabarcoding primers as capture probes to either isolate target DNA, or to remove nontarget DNA, prior to amplification by using biotinylated versions of universal metazoan and bacterial barcoding primers, namely for COI and bacterial 16S. Additionally, each step of the protocol was assessed by amplifying both COI and bacterial 16S to investigate the effect on the metazoan and bacterial communities. Bacterial abundance increased in response to the captures (COI library), while the quality of the captured DNA was improved. The metazoan-based probe captured bacterial DNA in a range that was also amplifiable with the 16S primers, demonstrating the ability of universal capture probes to isolate larger fragments of DNA from eDNA. This idea could be applied to metazoan metabarcoding, by using a truly conserved site without a high-level taxonomic resolution is targeted with aim to capture DNA spanning over a nearby the barcoding region, which can then be processed through a conventional metabarcoding by amplification protocol.

Identifier
Source https://data.blue-cloud.org/search-details?step=~012C71AEDBF825BCAA393A740CB829BCC64C9496BAA
Metadata Access https://data.blue-cloud.org/api/collections/C71AEDBF825BCAA393A740CB829BCC64C9496BAA
Provenance
Instrument Illumina NovaSeq 6000; ILLUMINA
Publisher Blue-Cloud Data Discovery & Access service; ELIXIR-ENA
Contributor UNIVERSITY COLLEGE DUBLIN
Publication Year 2024
OpenAccess true
Contact blue-cloud-support(at)maris.nl
Representation
Discipline Marine Science
Spatial Coverage (-87.495W, 15.864S, -87.495E, 15.864N)
Temporal Point 2019-07-01T00:00:00Z