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            <gco:CharacterString>Cold-water corals are conspicuous in the waters off Eastern Canada. Despite that, there are few DNA sequence records from specimens collected in the region available in GenBank, and not all species recorded in the region have sequence data regardless of geographic origin. This can limit the use of eDNA techniques to detect and identify corals. Our objective was to sequence and publish sequences for two octocoral DNA barcoding markers: CO1 and MutS. We sequenced and deposited 36 sequences to GenBank from 19 specimens representing three sea pen taxa (Octocorallia: Pennatuloidea): Distichoptilum gracile, Pennatula aculeata, and Protoptilum carpenteri. Identification of all specimens was confirmed by B. M. Neves before submission.  Specimens and DNA tissues were donated to the Canadian Museum of Nature, where they are currently stored. This publication is part 1 of a series of GenBank submissions by our lab.


Specimens were collected from across the Northwest Atlantic and originate from depths ranging between 200-1924 meters. Specimens were collected as part of research vessel multispecies trawl surveys or remotely operated vehicle (ROV ROPOS) surveys. DNA was isolated and purified using the QIAgen DNeasy Blood and Tissue kit, with an initial overnight incubation with Proteinase K. Two commonly used octocoral barcoding regions were amplified using previously described primers: 1) COII8068F (McFadden et al., 2004) and COIOCTR (France and Hoover, 2002) for the CO1 gene, and 2) ND42599F (France and Hoover, 2002) and mut3458R (Sánchez et al., 2003) for the MutS gene. Amplifications were conducted using 12.5 µl of Green DreamTaq Master Mix (Thermo Fisher Scientific), 1 µl of template DNA, 0.5 µl of each 10 µM forward and reverse primers, 0.5 µl of 10 µM reverse primer, and 10.5 µl of water. Thermocycling was run as follows: 3 min of initial denaturation at 95 °C, followed by 40 cycles at 95 °C for 30 s, 30 s at annealing temperature of 48 °C, then 65 s at an extension temperature of 72 °C, and a final elongation at 72 °C for 4 min. PCR products were cleaned using Agencourt AMPure XP Beads (Beckman Coulter) and sent to The Center for Advanced Genomics, Toronto, Canada for Sanger sequencing. Sequences were visualized and aligned using Geneious Prime 2022.0.2. Obtained sequences have been deposited in GenBank under accession numbers OQ569768- OQ569784 and OQ420359- OQ420377. 


This work was funded by Fisheries and Oceans Canada under an Enhanced Regional Capacity grant (2020-2021) and the Marine Conservation Targets (MCT) program (2021-2024), Newfoundland and Labrador Region.</gco:CharacterString>
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                  <gmd:LocalisedCharacterString locale="#fra">Les coraux d'eau froide sont communs dans les eaux au large de l'est du Canada. Malgré cela, peu de registres de séquences d’ADN de spécimens collectés dans la région sont disponibles dans GenBank, et toutes les espèces enregistrées dans la région ne disposent pas de données de séquence, quelle que soit leur origine géographique. Cela peut limiter l’utilisation de techniques comme l’ADN environnemental pour détecter et identifier les coraux. Notre objectif était de séquencer et de publier des séquences pour deux marqueurs de code-barres ADN pour les octocoraux : CO1 et MutS. Nous avons séquencé et déposé 36 séquences dans GenBank à partir de 19 spécimens représentant trois espèces de plume de mer (Octocorallia : Pennatuloidea) : Distichoptilum gracile, Pennatula aculeata et Protoptilum carpenteri. L'identification de tous les spécimens a été confirmée par B. M. Neves (MPO-TNL) avant la soumission.  Des spécimens et des tissus d'ADN ont été donnés au Musée canadien de la nature, où ils sont actuellement conservés. Cette publication représente la première partie d'une série de soumissions à GenBank par notre laboratoire


Les spécimens ont été collectés dans l’Atlantique nord-ouest et proviennent de profondeurs comprises entre 200 et 1924 mètres. Les spécimens ont été collectés dans le cadre de relevés au chalut multi-espèces menés par des navires de recherche ou de relevés effectués par des véhicules télécommandés (ROV ROPOS). L'ADN a été isolé et purifié à l'aide du kit QIAgen DNeasy Blood and Tissue, avec une incubation initiale d'une nuit avec la protéinase K. Deux régions de code-barres couramment utilisées pour les octocoraux ont été amplifiées à l'aide d'amorces décrites précédemment : 1) COII8068F (McFadden et al., 2004) et COIOCTR (France et Hoover, 2002) pour le gène CO1, et 2) ND42599F (France et Hoover, 2002) et mut3458R (Sánchez et al., 2003) pour le gène MutS. Les amplifications ont été réalisées en utilisant 12.5 µl de Green DreamTaq Master Mix (Thermo Fisher Scientific), 1 µl d'ADN matrice, 0.5 µl de chaque amorce sens et antisens à 10 µM, 0.5 µl d'amorce antisens à 10 µM et 10.5 µl d'eau. Le thermocyclage a été effectué comme suit : 3 minutes de dénaturation initiale à 95 °C, suivis de 40 cycles à 95 °C pendant 30 s, 30 s à une température de hybridation de 48 °C, puis 65 s à une température d'élongation de 72 °C, et un allongement final à 72 °C pendant 4 min. Les produits de PCR ont été purifiés à l'aide de l’Agencourt AMPure XP beads (Beckman Coulter) et envoyés au Centre for Applied Genomics, Toronto, Canada pour le séquençage Sanger. Les séquences ont été visualisées et alignées à l’aide de Geneious Prime 2022.0.2. Les séquences obtenues ont été déposées dans GenBank sous les numéros d'accès OQ569768-OQ569784 et OQ420359-OQ420377.


Ce projet a été financé par Pêches et Océans Canada dans le cadre d'une subvention pour le renforcement des capacités régionales (2020-2021) et du Programme des objectifs de conservation marine (MCT) (2021-2024), région de Terre-Neuve-et-Labrador.</gmd:LocalisedCharacterString>
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