Metadata: Identification_Information: Citation: Citation_Information: Originator: Christina A. Kellogg Originator: Julie J. Voelschow Publication_Date: 20210830 Title: Cold-water coral microbiomes (Acanthogorgia spp. Desmophyllum dianthus, and Lophelia pertusa) from the Gulf of Mexico and Atlantic Ocean off the southeast coast of the United States: raw sequencing data Geospatial_Data_Presentation_Form: FASTQ and tabular digital data Larger_Work_Citation: Citation_Information: Originator: Christina A. Kellogg Originator: Julie J. Voelschow Publication_Date: 20210830 Title: Cold-water Coral Microbiomes (Acanthogorgia spp., Desmophyllum dianthus, and Lophelia pertusa) from the Gulf of Mexico and Atlantic Ocean off the Southeast Coast of the United States—Raw Data Series_Information: Series_Name: U.S. Geological Survey Data Release Issue_Identification: doi:10.5066/P9Z1HPKR Publication_Information: Publication_Place: St. Petersburg, FL Publisher: U.S. Geological Survey Online_Linkage: https://doi.org/10.5066/P9Z1HPKR Description: Abstract: The files provided in this U.S. Geological Survey (USGS) data release (Kellogg and Voelschow, 2021) are the raw DNA sequence files referenced in the associated journal article (Kellogg and Pratte, 2021) entitled, “Unexpected diversity of Endozoicomonas in deep-sea corals.”. This dataset, PRJNA699458_16S-V3V4_raw_data_1.zip, represents the 16S rRNA gene amplicon surveys of 28 samples of deep-sea corals, including Acanthogorgia aspera (n=5), Acanthogorgia spissa (n=4), Desmophyllum dianthus (n=7), and Lophelia pertusa [Desmophyllum pertusum] (n=12), plus a kit extraction control blank. The sequencing targeted the V3-V4 variable region (primers 341F/806R) and was completed using an Illumina MiSeq sequencing system with version 2 chemistry to obtain paired-end reads. Purpose: During four research cruises conducted from 2013 to 2019, deep-sea coral samples were collected from various locations (nine sample sites, in total) in the Gulf of Mexico and Atlantic Ocean off the east coast of the United States for microbial analysis. Sample DNA extraction occurred at the USGS Coral Microbial Ecology Laboratory in St. Petersburg, Florida while polymerase chain reaction (PCR) amplification and sequencing were performed by RTSF Genomics Core at Michigan State University in East Lansing, Michigan. The 56 raw metagenomic data files associated with this data release have also been submitted to the National Center for Biotechnology Information (NCBI) Sequence Read Archive (SRA) under BioProject number PRJNA699458. Time_Period_of_Content: Time_Period_Information: Range_of_Dates/Times: Beginning_Date: 20130511 Ending_Date: 20190425 Currentness_Reference: ground condition Status: Progress: Complete Maintenance_and_Update_Frequency: None planned Spatial_Domain: Bounding_Coordinates: West_Bounding_Coordinate: -84.72679 East_Bounding_Coordinate: -74.51234 North_Bounding_Coordinate: 37.05002 South_Bounding_Coordinate: 26.20576 Description_of_Geographic_Extent: Gulf of Mexico and Atlantic Ocean Keywords: Theme: Theme_Keyword_Thesaurus: ISO 19115 Topic Category Theme_Keyword: biota Theme: Theme_Keyword_Thesaurus: USGS Metadata Identifier Theme_Keyword: USGS:3eb8d04f-e80f-48d6-8165-91810edced0c Theme: Theme_Keyword_Thesaurus: USGS Thesaurus Theme_Keyword: field sampling Theme_Keyword: laboratory methods Theme_Keyword: marine biology Theme_Keyword: microbiology Theme_Keyword: ecology Theme_Keyword: biodiversity Theme_Keyword: bacteria Place: Place_Keyword_Thesaurus: Thesaurus: Geographic Names Information System Place_Keyword: Gulf of Mexico Place_Keyword: Atlantic Ocean Access_Constraints: None. Please see 'Distribution Info' for details. Use_Constraints: Public domain data from the U.S. Government are freely redistributable with proper metadata and source attribution. The U.S. Geological Survey requests to be acknowledged as originator of these data in future products or derivative research. Point_of_Contact: Contact_Information: Contact_Person_Primary: Contact_Person: Christina A. Kellogg Contact_Organization: U.S. Geological Survey Contact_Address: Address_Type: mailing and physical Address: 600 4th Street South City: St. Petersburg State_or_Province: FL Postal_Code: 33701 Contact_Voice_Telephone: (727)-502-8128 Cross_Reference: Citation_Information: Originator: QIAGEN Publication_Date: 20200131 Title: DNEasy PowerBiofilm Kit Handbook Geospatial_Data_Presentation_Form: publication Online_Linkage: https://www.qiagen.com/us/resources/download.aspx?id=c5363245-cd9c-4314-b8fb-c4f8b4d669c4&lang=en Cross_Reference: Citation_Information: Originator: Christina A. Kellogg Originator: Zoe A. Pratt Publication_Date: 20210902 Title: Unexpected diversity of Endozoicomonas in deep-sea corals Geospatial_Data_Presentation_Form: publication Online_Linkage: https://doi.org/10.3354/meps13844 Data_Quality_Information: Attribute_Accuracy: Attribute_Accuracy_Report: No formal attribute accuracy tests were conducted. Logical_Consistency_Report: No formal logical consistency tests were conducted. Completeness_Report: Dataset is considered complete for the information presented, as described in the abstract. Lineage: Process_Step: Process_Description: Sample collection and preservation: Samples from Norfolk Canyon were collected in 2013 on the National Oceanic and Atmospheric Administration (NOAA) ship Ronald H. Brown using the remotely operated vehicle (ROV) Jason II (research cruise RB-13-03-HBH Deepwater Canyons). The samples acquired from west Florida sites Many Mounds and Okeanos Ridge were collected in 2017 on the NOAA ship Nancy Foster using the ROV Odysseus (NF1708 Southeast Florida Deep Coral Initiative). The 2018 and 2019 collections were part of NOAA's DeepSEARCH program and were conducted on the R/V Atlantis using the human occupied vehicle (HOV) Alvin (AT41) and the NOAA ship Ronald H. Brown using the ROV Jason II (RB1903), respectively. Subsamples of coral colonies were collected using the respective vehicle’s manipulator arm to remove a branch (or in the case of D. dianthus, the entire cup coral). Samples were placed into individual, thermally-insulated containers that had been precleaned (washed with freshwater, interiors wiped with 100% ethanol to remove any biofilms or particulates from prior collections), filled with freshwater, and sent down sealed. When opened to receive a coral collection, the freshwater was replaced by seawater local to the collection site due to density differences. The containers were then sealed at depth to prevent microbial contamination from other sample collections or passing through different water masses during vehicle recovery. At the conclusion of the dive, containers were brought aboard their respective vessel and into a cold room or laboratory and samples were removed using ethanol-sterilized forceps. All corals were lightly rinsed with sterile 1xPBS (phosphate-buffered saline) to remove any loosely associated surface microbes. For octocorals (A. aspera, A. spissa), branches that had not been in contact with the manipulator claw were cut off using ethanol-sterilized shears and transferred to sterile tubes. For the stony corals (D. dianthus, L. pertusa), samples were placed into sterile aluminum weigh boats and a flame-sterilized hammer was used to break open the calyces to expose polyp tissue, which was transferred to sterile tubes. All samples were preserved with RNAlater, stored for 24 hours at 4°C to allow the preservative to infiltrate the tissues, and then moved to -20 °C until processing. Subsamples of the corals collected for microbial analyses were shared with researchers conducting population genetics/taxonomy studies, allowing for accurate host identification of octocorals and the possibility of interpreting microbiome trends against coral genotypes. Process_Date: 20190425 Process_Step: Process_Description: DNA extraction: The DNA extraction process was performed on July 30th and July 312st, 2019, at the USGS Coral Microbial Ecology Laboratory. Just prior to extraction, coral samples were rinsed with 0.2 µm-filtered and autoclaved 1xPBS to remove RNAlater. Coral pieces were placed into sterile microcentrifuge tubes using flame-sterilized forceps, 2 ml of the sterile 1xPBS was added to the tube, which was then inverted 3 times and then centrifuged at 4 000 xg for 30 sec. The coral samples were then weighed using sterile technique and approximately 0.1–0.2 g per sample were used for extraction. Extractions were done in duplicate for each coral using Qiagen’s DNeasy PowerBiofilm kits (QIAGEN, 2020; catalog number 24000-50) and replicates were combined at the end. The manufacturer’s protocol was followed with the exception that a FastPrep was used on setting 5 (~3100 rpm) for homogenization in place of a PowerLyzer. Kit blank (extractions with no sample added) were processed at the same time as the samples. DNA was quantified using a Qubit, extractions diluted to approximately 30 ng/µl, and submitted for sequencing. Process_Date: 20190731 Process_Step: Process_Description: Sequencing: PCR amplification and sequencing were performed by Michigan State University Genomics Core. The V3-V4 regions of the 16S rRNA gene were amplified with the primers 341F (CCTACGGGAGGCAGCAG) and 806R (GGACTACHVGGGTWTCTAAT). One µl of genomic DNA was added to 7.5 µl of 2x Dream Taq Master Mix and 6.4 µl of a 0.5 µM primer mix. Amplification conditions were an initial melting period of 2 minutes at 95°C, followed by 30 cycles of 95°C for 40 seconds, 50°C for 30 seconds, and 72°C for 60 seconds, and a final anneal step at 72°C for 7 minutes. After PCR the output of all reactions was batch normalized using an Invitrogen SequalPrep DNA Normalization Plate, and all material recovered from the plate was pooled. The pooled material was cleaned up and concentrated using AmpureXP magnetic beads. The pool was quality controlled and quantified using a combination of Qubit dsDNA HS, Agilent 4200 TapeStation HS DNA1000 and Kapa Illumina Library Quantification qPCR assays. This pool was loaded onto an Illumina MiSeq v2 Standard flow cell and sequencing was carried out in a 2x250bp paired end format using a MiSeq v2 500 cycle reagent cartridge. Custom sequencing and index primers complementary to the 341F/806R sequences used for preparing the libraries were added to appropriate wells of the reagent cartridge. Base calling was done by Illumina Real Time Analysis (RTA) v1.18.54 and output of RTA was demultiplexed and converted to FastQ format with Illumina Bcl2fastq v2.19.1. Process_Date: 20201109 Entity_and_Attribute_Information: Overview_Description: Entity_and_Attribute_Overview: Please refer to the "README" file, README_PRJNA699458.txt, for detailed descriptions of the contents of the raw data files. Additional information is contained in the minimum information about a marker sequence (MIMARKS) compliant metadata, PRJNA699458_MIMARKS.xlsx, which is included in the data download files. Entity_and_Attribute_Detail_Citation: The entity and attribute information were generated by the individual and/or agency identified as the originator of the dataset. Please review the rest of the metadata record for additional details and information. Distribution_Information: Distributor: Contact_Information: Contact_Person_Primary: Contact_Person: Christina A. Kellogg Contact_Organization: St. Petersburg Coastal and Marine Science Center Contact_Address: Address_Type: mailing address Address: 600 4th Street South City: St. Petersburg State_or_Province: FL Postal_Code: 33701 Country: United States Contact_Voice_Telephone: (727) 502-8128 Contact_Electronic_Mail_Address: ckellogg@usgs.gov Distribution_Liability: Unless otherwise stated, all data, metadata and related materials are considered to satisfy the quality standards relative to the purpose for which the data were collected. Although these data and associated metadata have been reviewed for accuracy and completeness and approved for release by the U.S. Geological Survey (USGS), no warranty expressed or implied is made regarding the display or utility of the data on any other system or for general or scientific purposes, nor shall the act of distribution constitute any such warranty. Standard_Order_Process: Digital_Form: Digital_Transfer_Information: Format_Name: text, comma-delimited text, Microsoft Excel format File_Decompression_Technique: Winzip or 7zip Transfer_Size: 3050 Digital_Transfer_Option: Online_Option: Computer_Contact_Information: Network_Address: Network_Resource_Name: https://coastal.er.usgs.gov/data-release/doi-P9Z1HPKR/data/PRJNA699458_16S-V3V4_raw_data_1.zip Fees: None Metadata_Reference_Information: Metadata_Date: 20211109 Metadata_Contact: Contact_Information: Contact_Person_Primary: Contact_Person: Christina A. Kellogg Contact_Organization: St. Petersburg Coastal and Marine Science Center Contact_Address: Address_Type: mailing address Address: 600 4th Street South City: St. Petersburg State_or_Province: FL Postal_Code: 33701 Contact_Voice_Telephone: (727) 502-8128 Contact_Electronic_Mail_Address: ckellogg@usgs.gov Metadata_Standard_Name: FGDC Biological Data Profile of the Content Standard for Digital Geospatial Metadata Metadata_Standard_Version: FGDC-STD-001.1-1999