9806a672-b859-4175-8432-f805b5ce8f30 https://ipt-obis.gbif.us/resource?r=noaa-oer-okeanos-ex2301 noaa-oer-okeanos-ex2301 Clement Coclet NOAA Atlantic Oceanographic and Meteorological Laboratory Research Scientist II

4301 Rickenbacker Cswy Miami Florida 33149 US

clement.coclet@noaa.gov 0000-0002-6672-148X Luke Thompson NOAA Atlantic Oceanographic and Meteorological Laboratory Research Professor

4301 Rickenbacker Cswy Miami Florida 33149 US

luke.thompson@noaa.gov 0000-0002-3911-1280 Nicole Miller NOAA Ocean Exploration NOAA Omics Coordinator

1315 East-West Hwy Silver Spring Maryland 20910 US

nicole.miller@noaa.gov 0009-0002-8148-6323 Ashley Marranzino NOAA Ocean Exploration Project Scientist

1315 East-West Hwy Silver Spring Maryland 20910 US

ashley.marranzino@noaa.gov 0000-0002-6451-1139 Jennifer Green NOAA Ocean Exploration Research Associate III

1021 Balch Blvd Ste 1003 Stennis Space Center Mississippi 39529 US

jennifer.green@noaa.gov Logan Kline NOAA Ocean Exploration Operations Support Analyst

1315 East-West Hwy Silver Spring Maryland 20910 US

logan.kline@noaa.gov 0000-0003-0257-9397 Allen Collins NOAA National Systematics Laboratory Director and Research Zoologist

MRC 163 Washington District of Columbia 20013-7012 US

allen.collins@noaa.gov 0000-0002-3664-9691 Steven Auscavitch Smithsonian National Museum of Natural History Genomics Specialist

MRC 163 Washington District of Columbia 20013-7012 US

auscavitchs@si.edu 0000-0001-5777-4814 2026-05-06 eng Environmental DNA (eDNA) collected by NOAA Ship Okeanos Explorer during expedition EX2301: 2023 Shakedown + EXPRESS West Coast Exploration (April 13-27, 2023), Portland, Oregon to Seattle, Washington. eDNA samples (48) collected by ROV and metabarcoded using 12S, 18S (V9), and COI markers. marine biome ENVO marine epipelagic zone ENVO marine mesopelagic zone ENVO marine bathypelagic zone ENVO seawater ENVO Pacific Ocean N/A 18S rRNA (SSU eukaryote) N/A 12S rRNA (SSU mitochondria) N/A COI (cytochrome oxidase I) N/A eDNA ASFA metabarcoding ASFA marine biodiversity ASFA environmental DNA GCMD Occurrence GBIF Dataset Type Vocabulary: http://rs.gbif.org/vocabulary/gbif/dataset_type_2015-07-10.xml To the extent possible under law, the publisher has waived all rights to these data and has dedicated them to the Public Domain (CC0 1.0). Users may copy, modify, distribute and use the work, including for commercial purposes, without restriction. Creative Commons Zero v1.0 Universal https://spdx.org/licenses/CC0-1.0.html CC0-1.0 https://www.ncei.noaa.gov/waf/okeanos-rov-cruises/ex2301/ https://ipt-obis.gbif.us/archive.do?r=noaa-oer-okeanos-ex2301 Northeast Pacific (U.S. West Coast continental margin, northern California to Washington, ~40–48°N, ~125–126°W) -126.126 -124.755 48.171 40.133 2023-04-15 2023-04-26 Marine organisms identified through eDNA metabarcoding kingdom Animalia kingdom Bacteria kingdom Chromista kingdom Fungi kingdom Plantae kingdom Protozoa This dataset serves two purposes. First, to advance alignment among researchers and stakeholders on making NOAA Ocean Exploration eDNA data publicly available and interpretable, and to identify and address challenges in achieving that goal. Second, to characterize biodiversity in sampled ocean environments using eDNA metabarcoding data from NOAA Ship Okeanos Explorer expeditions, with emphasis on deep-sea communities in the abyssal zone (depths below 4,000 m). asNeeded Luke Thompson NOAA Atlantic Oceanographic and Meteorological Laboratory Research Professor

4301 Rickenbacker Cswy Miami Florida 33149 US

luke.thompson@noaa.gov https://www.linkedin.com/in/luke-thompson-bb20074/ 0000-0002-3911-1280 DNA was extracted from seawater filters, amplified by PCR for 18S, COI, and 12S metabarcoding markers, sequenced on Illumina, and analyzed with the Tourmaline 2 pipeline. Field sampling design and detailed procedures are described under sampling; bioinformatic QC and filtering are described under quality control. Northeast Pacific (U.S. West Coast continental margin, northern California to Washington, ~40–48°N, ~125–126°W) from 2023-04-15 to 2023-04-26. Seawater samples were collected aboard NOAA Ship Okeanos Explorer during ROV dives and CTD casts. Water was drawn from Niskin bottles mounted on either the Deep Discoverer ROV or a CTD rosette. Standard collection volumes were 1.7 L for water samples and 2.0 L for distilled water blanks. Samples were filtered through 0.45 um or 0.22 um membrane filters. Sample and environmental metadata - including dive or CTD cast ID, date, time, location, in-situ measurements (salinity, temperature, dissolved oxygen), and estimated bottom depth - were recorded electronically and transferred to the Sample Operator User Portal (SOUP). Laboratory procedures followed documented protocols (https://github.com/aomlomics/protocols). DNA was extracted from filters and amplified by PCR targeting three metabarcoding markers: 18S rRNA (protists/eukaryotes), COI (metazoa), and 12S rRNA (fish). Sequencing was performed on the Illumina NextSeq platform at the NOAA National Systematics Laboratory. Bioinformatic processing was performed on the Northern Gulf Institute's Orion computing cluster using the Tourmaline 2 pipeline. Steps included adapter trimming and quality filtering (Cutadapt), denoising and representative sequence determination (DADA2), and taxonomic assignment using a naive Bayes classifier and consensus VSEARCH (QIIME 2). Raw sequence data in FASTQ format were submitted to the NCBI Sequence Read Archive (SRA). FAIRe-formatted sample metadata were submitted to NCBI BioSample. Taxonomic occurrence data were formatted for submission to OBIS. Quality control measures were implemented across all stages of the workflow. During sample collection, laboratory errors and deviations from standard operating procedures (NOAA Ocean Exploration eDNA Sample Workflow, Revision 2.3) were recorded per sample in a physical notebook and transferred to the Sample Operator User Portal (SOUP). Negative controls (distilled water blanks, extraction blanks, PCR blanks, and sequencing blanks) were included throughout sampling and processing to monitor for contamination. Control samples were excluded from the final published dataset but are archived in NCBI SRA and BioSample records for transparency. Raw sequence data were processed using the Tourmaline 2 pipeline. Low-quality bases and adapters were trimmed using Cutadapt, and sequences were denoised using DADA2 to generate representative sequences. Taxonomic assignments were made using a naive Bayes classifier and consensus VSEARCH, with confidence thresholds applied to ensure reliable identifications. Post-assignment filters were applied to remove sequences associated with domestic or human taxa (Bovidae, Suidae, Felidae, Canidae, Primates, Homo sapiens), unresolved assignments (Unassigned), and terrestrial organisms as flagged by WoRMS habitat classifications. Luke Thompson 0000-0002-3911-1280 processor Clement Coclet 0000-0002-6672-148X processor Nicole Miller 0009-0002-8148-6323 curator Ashley Marranzino 0000-0002-6451-1139 curator Jennifer Green curator Logan Kline 0000-0003-0257-9397 originator Allen Collins 0000-0002-3664-9691 reviewer Steven Auscavitch 0000-0001-5777-4814 originator Sammy Harding 0009-0008-8885-6140 curator Bayden Willms 0009-0003-2751-7317 programmer Katherine Silliman 0000-0001-5964-3965 publisher The mission of NOAA Ocean Exploration is to explore the ocean for national benefit, including discovery and characterization of deep-sea environments and biodiversity. This project provides preliminary environmental DNA metabarcoding data from plankton, metazoa, cnidaria, and fish collected aboard NOAA Ship Okeanos Explorer, operated by NOAA Ocean Exploration. Sample archiving and management, wet lab processing, and sequencing was conducted by the Smithsonian National Museum of Natural History and the NOAA National Systematics Laboratory. DNA sequence analysis was performed by the NOAA Atlantic Oceanographic and Meteorological Laboratory. "A Comprehensive Bioinformatics Platform for Marine eDNA and Microbiome Data" (Grant ID NA21OAR4320190-T3-01S040, NOAA Ocean Exploration) NOAA Ocean Exploration NA21OAR4320190-T3-01S040 Atlantic (SE U.S., Gulf of America, Puerto Rico/Puerto Rico Trench, Azores) and Pacific (U.S. West Coast) from 2021-10-27 to 2023-04-26. To characterize biodiversity in sampled ocean environments using eDNA metabarcoding data from NOAA Ship Okeanos Explorer expeditions, with emphasis on deep-sea communities in the abyssal zone (depths below 4,000 m). 2026-04-13T20:08:09.322+00:00 dataset Coclet C, Thompson L, Miller N, Marranzino A, Green J, Kline L, Collins A, Auscavitch S (2026). NOAA Ocean Exploration seawater eDNA metabarcoding (EX2301). Version 1.4. NOAA Ocean Exploration. Occurrence dataset. https://ipt-obis.gbif.us/resource?r=noaa-oer-okeanos-ex2301&v=1.4 9806a672-b859-4175-8432-f805b5ce8f30/v1.4.xml