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            <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/dataset/998297.rdf" xlink:actuate="onRequest">Decorator worm Diopatra cuprea single nucleotide polymorphism (SNP) genotypes from collections in the Gulf of Mexico and eastern United States estuaries from 2009 to 2022</gmx:Anchor>
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            <gco:CharacterString>Cite this dataset as: Sotka, E. (2026) Decorator worm Diopatra cuprea single nucleotide polymorphism (SNP) genotypes from collections in the Gulf of Mexico and eastern United States estuaries from 2009 to 2022. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2026-05-08 [if applicable, indicate subset used]. http://lod.bco-dmo.org/id/dataset/998297 [access date]</gco:CharacterString>
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        <gco:CharacterString>Dataset Description: &amp;lt;p&amp;gt;See &amp;quot;Related Datasets&amp;quot; for&amp;amp;nbsp;sanger-sequenced a mitochondrial locus (COI) metadata and accession information at NCBI's Genbank database.&amp;lt;/p&amp;gt; Methods and Sampling: &amp;lt;p&amp;gt;Note: This metadata section includes description of a related dataset (sanger-sequenced a mitochondrial locus (COI) that was generated as part of this study. See &amp;quot;Related Datasets&amp;quot; section for metadata and NCBI Genbank accession identifiers in that dataset.&amp;amp;nbsp;&amp;lt;br /&amp;gt;
&amp;lt;br /&amp;gt;
&amp;lt;em&amp;gt;Diopatra cuprea&amp;lt;/em&amp;gt; were sampled from 19 locations on the United States east coast, from Duxbury MA (the genus’ northern limit) to St. Teresa Beach FL. Tubes were excavated with a shovel, the worm removed, and antennae clipped off and preserved in 95% ethanol for later DNA extraction. or 2022 samples, twenty-five randomly selected individuals from each population were extracted for DNA, with the exception of Broad River Estuary populations from which 20 and 22 samples were collected. Approximately 25&amp;amp;nbsp;mg of tissue wet weight was&amp;amp;nbsp;rinsed of ethanol with deionized water and extracted with the Qiagen DNeasy Blood and Tissue Kit (Qiagen, Valencia, CA, USA) following the manufacturer’s extraction protocol. Extractions were screened with 1.5% agarose gels and DNA was quantified using a NanoDrop 200 spectrophotometer (Thermo Scientific, Waltham, MA, USA) to obtain concentration values and purity.&amp;amp;nbsp;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;A portion of the COI gene was then PCR amplified using the protocol in Berke, et al. (2010, doi:10.1111/j.1466-8238.2009.00509.x). These were cleaned with an EXO-SAP-IT protocol and sent for Sanger sequencing with these same primers at a private company.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;Double digest restriction-associated DNA sequencing (or ddRADSeq) library was prepared on 312 samples following the protocol in Parchman et al. (2012,doi: 10.1111/j.1365-294X.2012.05513.x).&amp;amp;nbsp;Briefly, we digested gDNA with two restriction enzymes, EcoRI and MseI, and ligated adaptors containing unique 8 to 10&amp;amp;nbsp;bp barcodes to the digested DNA of each individual. The products were then PCR amplified in two independent reactions with standard Illumina primers. All amplicons were pooled and shipped to the University of Texas Genomic Sequencing and Analysis Facility or the Tufts University Core Facility, which used Pippin Prep&amp;lt;sup&amp;gt;®&amp;lt;/sup&amp;gt; to isolate the 300–450&amp;amp;nbsp;bp fraction. This fraction was then single-read sequenced (100 basepairs) with Illumina HiSeq 4000 machine. FASTQ sequences were demultiplexed using custom Unix code. 510&amp;amp;nbsp;million reads contained barcode sequence (range = 39 to 6.8&amp;amp;nbsp;million (M) reads per sample; mean = 1.6&amp;amp;nbsp;M reads), and 234 individuals had at least 200&amp;amp;nbsp;K reads that were analyzed.&amp;lt;br /&amp;gt;
&amp;lt;br /&amp;gt;
FASTQ files were uploaded to the NCBI's Sequence Read Archive (SRA) that were generated as described above.&amp;amp;nbsp; See the &amp;quot;Related Datasets&amp;quot; section&amp;amp;nbsp;for SRA metadata and accessions.&amp;amp;nbsp;&amp;lt;br /&amp;gt;
&amp;lt;br /&amp;gt;
Organism identifier (Life Science Identifier (LSID)):&amp;lt;br /&amp;gt;
Diopatra cuprea, Bosc, 1802, LSID(urn:lsid:marinespecies.org:taxname:157339)&amp;lt;/p&amp;gt;</gco:CharacterString>
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        <gmx:Anchor xlink:href="http://lod.bco-dmo.org/id/award/820199.rdf" xlink:title="OCE-1924599" xlink:actuate="onRequest">Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1924599 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1924599</gmx:Anchor>
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&lt;p&gt;During the 20th century, the Pacific oyster &lt;em&gt;Crassostrea gigas&lt;/em&gt; was deliberately introduced from its native range of coastal Asia to the estuaries of six continents. While the introduced Pacific oysters are widely aquacultured and thus can generate local economic wealth, they sometimes outcompete native oysters, and can carry microbial, animal and plant hitchhikers that negatively impact local economies and the ecological functioning of local estuaries. This study comprehensively assesses the pathways and sources of Pacific oyster introductions using a worldwide, population genetic survey. Simultaneously, the study also assesses the pathways and source of one hitchhiking protist (&lt;em&gt;Haplosporidium nelsoni&lt;/em&gt;) that causes the disease MSX (multinucleated sphere X) in the Virginia oyster (&lt;em&gt;Crassostrea virginica&lt;/em&gt;) along the eastern seaboard of the United States. One goal of this research is to generate management strategies that combat the negative impacts of the Pacific oyster and its associated invaders, and minimize future invasions. A second goal is to minimize some uncertainty about the population biology of the devastating Haplosporidium parasite, and thus, increase confidence of policy makers who are managing shellfish health, restoration and commerce. By quantifying the pathways and sources of &lt;em&gt;C. gigas&lt;/em&gt;, this project may inform strategies to combat negative impacts of &lt;em&gt;C. gigas&lt;/em&gt; and its associated invaders, as well as minimize future invasions. Moreover, quantifying dispersal within and among populations of &lt;em&gt;H. nelsoni&lt;/em&gt; along the US East Coast will provide perspective on the effectiveness of regional biosecurity measures in preventing the ongoing dispersal of this destructive pathogen via aquaculture. In addition, the project lends itself well to programs that foster critical thinking and research experience among both undergraduate and K-12 students. The project provides opportunities for 6-9 undergraduates to perform research, includes a 2-day workshop on bioinformatics for the wider undergraduate community, and facilitates ongoing opportunities for K-12 students to participate in citizen-science research.&lt;/p&gt;
&lt;p&gt;There is a wealth of information on the source, pathways and vectors of &lt;em&gt;C. gigas&lt;/em&gt; based largely on historical documents but no study has comprehensively tested whether these historical accounts are correct using a worldwide, population genetic survey. Using &amp;gt;14K single-nucleotide polymorphisms (SNPs) from 41 populations across five continents a high level of spatial genetic differentiation was found within the native range and differences in source populations among non-native regions. Preliminary genetic data indicated that the parasitic protist, &lt;em&gt;Haplosporidium nelsoni &lt;/em&gt;arrived with C. gigas imports to the US Atlantic coastline and then infected the native C. virginica, however the native source populations, the pathways and vector from which &lt;em&gt;H. nelsoni &lt;/em&gt;arrived remain unknown. This project couples high-throughput sequencing technologies and Approximate Bayesian Computing (ABC)-based models to answer the following: What are the population genomic patterns among &lt;em&gt;C. gigas&lt;/em&gt; from native and non-native regions? What are the population genomic patterns of &lt;em&gt;Haplosporidium nelsoni &lt;/em&gt;among Asian and North American Crassostrea gigas and eastern North American &lt;em&gt;C. virginica&lt;/em&gt;? What were the source populations and invasion pathways of &lt;em&gt;C. gigas&lt;/em&gt; and &lt;em&gt;H. nelsoni&lt;/em&gt;? Identifying source locations, pathways and vectors of introduction of C. gigas will provide researchers with a null-model of invasion history for dozens of other non-native species that were transported with &lt;em&gt;C. gigas&lt;/em&gt;. Currently, there are no verified 'vector maps' for historical shipments of &lt;em&gt;C. gigas&lt;/em&gt; that are similar to those generated from modern-day or historical shipping records.&lt;/p&gt;
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http://lod.bco-dmo.org/id/dataset-parameter/999358.rdf
	Name: sample_id
	Units: unitless
	Description: &lt;p&gt;individual sample ID&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999359.rdf
	Name: pop_defined
	Units: unitless
	Description: &lt;p&gt;Popular name of site&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999360.rdf
	Name: geographic_location
	Units: unitless
	Description: &lt;p&gt;Sample Site&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999361.rdf
	Name: state_name
	Units: unitless
	Description: &lt;p&gt;State name (USA)&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999362.rdf
	Name: estuary_local
	Units: unitless
	Description: &lt;p&gt;Coastal vs Inland&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999363.rdf
	Name: mt_haplotype
	Units: unitless
	Description: &lt;p&gt;mtDNA haplotype from Sotka et al. 2023 doi.org/10.3390/biology12040521&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999364.rdf
	Name: mt_hap_group
	Units: unitless
	Description: &lt;p&gt;mtDNA haplotype from Sotka et al. 2023 doi.org/10.3390/biology12040521&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999365.rdf
	Name: mt_hap_subgroup
	Units: unitless
	Description: &lt;p&gt;mtDNA haplotype from Sotka et al. 2023 doi.org/10.3390/biology12040521&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999366.rdf
	Name: collector
	Units: unitless
	Description: &lt;p&gt;Person&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999367.rdf
	Name: SRA_Run
	Units: unitless
	Description: &lt;p&gt;National Center for Biotechnology Information (NCBI) Run accession in the Sequence Read Archive (SRA).&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999368.rdf
	Name: SRA_Study
	Units: unitless
	Description: &lt;p&gt;National Center for Biotechnology Information (NCBI) study accession in the Sequence Read Archive (SRA).&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999369.rdf
	Name: version
	Units: unitless
	Description: &lt;p&gt;SRA accession version number&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999370.rdf
	Name: BioSample
	Units: unitless
	Description: &lt;p&gt;National Center for Biotechnology Information (NCBI) BioSample accession&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999371.rdf
	Name: site_lat
	Units: decimal degrees
	Description: &lt;p&gt;Site latitude&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999373.rdf
	Name: site_lon
	Units: decimal degrees
	Description: &lt;p&gt;Site longitude&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999375.rdf
	Name: sites_collector
	Units: unitless
	Description: &lt;p&gt;co-author that collected samples&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999376.rdf
	Name: Assay_Type
	Units: unitless
	Description: &lt;p&gt;Assay Type&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999377.rdf
	Name: AvgSpotLen
	Units: count
	Description: &lt;p&gt;Average Spot Length (SRA metadata from NCBI)&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999378.rdf
	Name: Bases
	Units: count
	Description: &lt;p&gt;Bases&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999379.rdf
	Name: BioProject
	Units: unitless
	Description: &lt;p&gt;National Center for Biotechnology Information (NCBI) BioProject identifier.&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999380.rdf
	Name: BioSampleModel
	Units: unitless
	Description: &lt;p&gt;BioSample model (e.g. Invertebrate)&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999381.rdf
	Name: Bytes
	Units: bytes
	Description: &lt;p&gt;Size in bytes&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999382.rdf
	Name: Experiment
	Units: unitless
	Description: &lt;p&gt;National Center for Biotechnology Information (NCBI) Experiment accession in the Sequence Read Archive (SRA).&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999383.rdf
	Name: Instrument
	Units: unitless
	Description: &lt;p&gt;Instrument name&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999384.rdf
	Name: isolate
	Units: unitless
	Description: &lt;p&gt;Isolate&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999385.rdf
	Name: isolation_source
	Units: unitless
	Description: &lt;p&gt;isolation source&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999386.rdf
	Name: Library_Name
	Units: unitless
	Description: &lt;p&gt;Library Name&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999387.rdf
	Name: LibraryLayout
	Units: unitless
	Description: &lt;p&gt;Library Layout&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999388.rdf
	Name: LibrarySelection
	Units: unitless
	Description: &lt;p&gt;Library Selection&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999389.rdf
	Name: LibrarySource
	Units: unitless
	Description: &lt;p&gt;Library Source&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999390.rdf
	Name: Organism
	Units: unitless
	Description: &lt;p&gt;organism sampled (Diopatra cuprea)&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999391.rdf
	Name: Platform
	Units: unitless
	Description: &lt;p&gt;platform&lt;/p&gt; 
http://lod.bco-dmo.org/id/dataset-parameter/999392.rdf
	Name: tissue
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	Description: &lt;p&gt;tissue sample location&lt;/p&gt; 
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&amp;lt;br /&amp;gt;
&amp;lt;em&amp;gt;Diopatra cuprea&amp;lt;/em&amp;gt; were sampled from 19 locations on the United States east coast, from Duxbury MA (the genus’ northern limit) to St. Teresa Beach FL. Tubes were excavated with a shovel, the worm removed, and antennae clipped off and preserved in 95% ethanol for later DNA extraction. or 2022 samples, twenty-five randomly selected individuals from each population were extracted for DNA, with the exception of Broad River Estuary populations from which 20 and 22 samples were collected. Approximately 25&amp;amp;nbsp;mg of tissue wet weight was&amp;amp;nbsp;rinsed of ethanol with deionized water and extracted with the Qiagen DNeasy Blood and Tissue Kit (Qiagen, Valencia, CA, USA) following the manufacturer’s extraction protocol. Extractions were screened with 1.5% agarose gels and DNA was quantified using a NanoDrop 200 spectrophotometer (Thermo Scientific, Waltham, MA, USA) to obtain concentration values and purity.&amp;amp;nbsp;&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;A portion of the COI gene was then PCR amplified using the protocol in Berke, et al. (2010, doi:10.1111/j.1466-8238.2009.00509.x). These were cleaned with an EXO-SAP-IT protocol and sent for Sanger sequencing with these same primers at a private company.&amp;lt;/p&amp;gt;

&amp;lt;p&amp;gt;Double digest restriction-associated DNA sequencing (or ddRADSeq) library was prepared on 312 samples following the protocol in Parchman et al. (2012,doi: 10.1111/j.1365-294X.2012.05513.x).&amp;amp;nbsp;Briefly, we digested gDNA with two restriction enzymes, EcoRI and MseI, and ligated adaptors containing unique 8 to 10&amp;amp;nbsp;bp barcodes to the digested DNA of each individual. The products were then PCR amplified in two independent reactions with standard Illumina primers. All amplicons were pooled and shipped to the University of Texas Genomic Sequencing and Analysis Facility or the Tufts University Core Facility, which used Pippin Prep&amp;lt;sup&amp;gt;®&amp;lt;/sup&amp;gt; to isolate the 300–450&amp;amp;nbsp;bp fraction. This fraction was then single-read sequenced (100 basepairs) with Illumina HiSeq 4000 machine. FASTQ sequences were demultiplexed using custom Unix code. 510&amp;amp;nbsp;million reads contained barcode sequence (range = 39 to 6.8&amp;amp;nbsp;million (M) reads per sample; mean = 1.6&amp;amp;nbsp;M reads), and 234 individuals had at least 200&amp;amp;nbsp;K reads that were analyzed.&amp;lt;br /&amp;gt;
&amp;lt;br /&amp;gt;
FASTQ files were uploaded to the NCBI's Sequence Read Archive (SRA) that were generated as described above.&amp;amp;nbsp; See the &amp;quot;Related Datasets&amp;quot; section&amp;amp;nbsp;for SRA metadata and accessions.&amp;amp;nbsp;&amp;lt;br /&amp;gt;
&amp;lt;br /&amp;gt;
Organism identifier (Life Science Identifier (LSID)):&amp;lt;br /&amp;gt;
Diopatra cuprea, Bosc, 1802, LSID(urn:lsid:marinespecies.org:taxname:157339)&amp;lt;/p&amp;gt;</gco:CharacterString>
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              <gmd:source>
                <gmd:LI_Source>
                  <gmd:sourceCitation>
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                      <gmd:title>
                        <gco:CharacterString>Specified by the Principal Investigator(s)</gco:CharacterString>
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                  <gco:CharacterString>- Loaded TSV file SRAattributes+meta.txt as table 998297_v1_diopatra-sra-metadata, with empty string, &amp;quot;nd&amp;quot;, and &amp;quot;NA&amp;quot; treated as missing values
- Renamed column: accession to BioSample
- Applied metadata (descriptions, standard name IDs, units) to 14 columns in 998297_v1_diopatra-sra-metadata including sample_ID, geographic.location, sites.Lat, sites.Lon, mtHaplotype, and related fields
- Renamed 13 columns in 998297_v1_diopatra-sra-metadata: sample_ID to sample_id, Pop.Defined to pop_defined, geographic.location to geographic_location, State.Name to state_name, EstuaryLocal to estuary_local, mtHaplotype to mt_haplotype, mtHap.Group to mt_hap_group, mtHap.Subgroup to mt_hap_subgroup, Year.Collected to year_collected, Collector to collector, sites.Lat to site_lat, sites.Lon to site_lon, sites.Collector to sites_collector
- Negated site_lon values (multiplied by -1) using a computed field mathematical operation (longitudes W are negative)
- Rounded site_lat and site_lon to 5 decimal places with trailing zeros preserved
- Loaded CSV file SraRunTable_PRJNA1103840.csv (output from NCBI Run Selector Tool) using filename as table name, with empty string and &amp;quot;nd&amp;quot; treated as missing values
- Joined SRA metadata from the NCBI Run selector into provided metadata table using BioSample as the join key.  Tables both were checked first that they were unique by BioSample.  Added SRA metadata not present in the attributes file including Assay Type, AvgSpotLen, Bases, BioProject, BioSampleModel, Bytes, Experiment, Instrument, Library Name, LibraryLayout, LibrarySelection, LibrarySource, Organism, Platform, SRA Run, SRA Study, isolate, isolation_source, tissue, and version
- Reordered 34 columns, placing year_collected and sample_id first, followed by sample metadata, SRA identifiers, and sequencing metadata
- Renamed columns: Assay Type to Assay_Type, Library Name to Library_Name
- Applied updated metadata (descriptions, standard name IDs, units) to all 34 columns
- Set final data types for columns
- Output written to 998297_v1_diopatra-sra-metadata.csv</gco:CharacterString>
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