metabarcoding
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The ABYSS project aims at describing deep-sea benthic biodiversity spanning several branches of the tree of life with eDNA metabarcoding tools. To accommodate both micro- and macro biologists, we designed a bioinformatic pipeline based on Illumina read correction with Dada2 allowing analysing metabarcodes from prokaryotic and eukaryotic life compartments.
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In October 2019 we chose 15 sites from the 2019 EVHOE survey for environmental DNA (eDNA) sampling. The French international EVHOE bottom trawl survey is carried out annually during autumn in the BoB to monitor demersal fish resources. At each site, we sampled seawater using Niskin bottles deployed with a circular rosette. There were nine bottles on the rosette, each of them able to hold ∼5 l of water. At each site, we first cleaned the circular rosette and bottles with freshwater, then lowered the rosette (with bottles open) to 5 m above the sea bottom, and finally closed the bottles remotely from the boat. The 45 l of sampled water was transferred to four disposable and sterilized plastic bags of 11.25 l each to perform the filtration on-board in a laboratory dedicated to the processing of eDNA samples. To speed up the filtration process, we used two identical filtration devices, each composed of an Athena® peristaltic pump (Proactive Environmental Products LLC, Bradenton, Florida, USA; nominal flow of 1.0 l min–1 ), a VigiDNA 0.20 μm filtration capsule (SPYGEN, le Bourget du Lac, France), and disposable sterile tubing. Each filtration device filtered the water contained in two plastic bags (22.5 l), which represent two replicates per sampling site. We followed a rigorous protocol to avoid contamination during fieldwork, using disposable gloves and single-use filtration equipment and plastic bags to process each water sample. At the end of each filtration, we emptied the water inside the capsule that we replaced by 80 ml of CL1 conservation buffer and stored the samples at room temperature following the specifications of the manufacturer (SPYGEN, Le Bourget du Lac, France). We processed the eDNA capsules at SPYGEN, following the protocol proposed by Polanco-Fernández et al., (2020). Half of the extracted DNA was processed by Sinsoma using newly developped ddPCR assays for European seabass (Dicentrachus labrax), European hake (Merluccius merluccius) and blackspot seabream (Pagellus bogaraveo). The other half of the extracted DNA was analysed using metabarcoding with teleo primer. The raw metabarcoding data set is available at https://www.doi.org/10.16904/envidat.442 Bottom trawling using a GOV trawl was carried out before or after water sampling. The catch was sorted by species and catches in numbers and weight were recorded. No blackspot seabream individuals were caught. Data content: * ddPCR/: contains the ddPCR counts and DNA concentrations for each sample and species. * SampleInfo/: contains the filter volume for each eDNA sample. * StationInfo/: contains metadata related to the data collected in the field for each filter. * Metabarcoding/: contains metabarcoding results for teleoprimer. * Trawldata/: contains catch data in numbers and weight (kg).
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Arctic marine protist communities have been understudied due to challenging sampling conditions, in particular during winter and in deep waters. The aim of this study was to improve our knowledge on Arctic protist diversity through the year, both in the epipelagic (< 200 m depth) and mesopelagic zones (200-1000 m depth). Sampling campaigns were performed in 2014, during five different months, to capture the various phases of the Arctic primary production: January (winter), March (pre-bloom), May (spring bloom), August (post-bloom) and November (early winter). The cruises were undertaken west and north of the Svalbard archipelago, where warmer Atlantic waters from the West Spitsbergen Current meets cold Arctic waters from the Arctic Ocean. From each cruise, station, and depth, 50 L of sea water were collected and the plankton was size-fractionated by serial filtration into four size fractions between 0.45-200 µm, representing the picoplankton, nanoplankton and microplankton. In addition vertical net hauls were taken from 50 m depth to the surface at selected stations. From the plankton samples DNA was extracted, the V4 region of the 18S rRNA-gene was amplified by PCR with universal eukaryote primers and the amplicons were sequenced by Illumina high-throughput sequencing. Sequences were clustered into Amplicon Sequence Variants (ASVs), representing protist genotypes, with the dada2 pipeline. Taxonomic classification was made against the curated Protist Ribosomal Reference database (PR2). Here we present the ASV table, metadata for the sequenced size-fractionated plankton samples, and environmental data from the corresponding water masses. Fastq-files with raw reads are available from the European Nucleotide Archive, under project accession number PRJEB40133.
Catalogue PIGMA