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    Populating NCBI template for submissions using BioNumerics

    Ruth Timme, Maria Balkey, Julie Haendiges, Tina Lusk Pfefer, Brian Sauders

    Published: 2023-12-21 DOI: 10.17504/protocols.io.36wgq3ejylk5/v2
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    Abstract

    PURPOSE: to define the standard operating procedure for collecting isolate metadata using BioNumerics for submission of food/environmental isolates to NCBI.

    SCOPE: to provide a standardized procedure to collect isolate metadata using BioNumerics for submission of food/environmental isolates to NCBI.

    RESPONSIBILITIES - SOP Responsible Officials: Ruth Timme, Maria Balkey

    The GenomeTrakr Network Management will be responsible to monitor GenomeTrakr submissions processed through Bionumerics and ensure that all GT labs are familiar with the mandatory metadata fields required for submission of GenomeTrakr sequencing records to NCBI.

    V3: Dropdown menus from controlled vocabulary added to the ncbi_update submission sheet

    Steps

    1.

    Metadata SampleSheet preparation

    Before uploading your sequencing run or linking NCBI sequencing records at the BioNumerics platform make sure to fill out the metadata spreadsheet form.

    Please download the template and guidelines included in the file GT_BioNumerics_spreadsheet_v3.xlsx

    Create the fields NCBI_bioproject, Attribute_package, Organism_name, NCBI_LabID, SourceCountryState, Latitude_longitude, Reference_material, Culture_collection, ProjectName, SequencedBy or Description if they are not in the BioNumerics interface and are needed to process the metadata for your isolates.

    Once you have filled out the template information, save the template sheet as .csv and import the metadata to BioNumerics.

    2.

    NCBI Submission Settings (Manage submission template)

    Create the NCBI metadata template in BioNumerics following PulseNet instructions making sure fields are populated according to GT requirements which are described in the following steps.

    2.1.

    BioProject and Organization: GenomeTrakr labs by submitting independently become owners of their data and are responsible for managing individual bioprojects for each sequenced organism. The term 'field content ' field content ' denotes that the template value e.g. BioProject accession is mapping to the field in BioNumerics e.g. NCBI_bioproject.

    Fig 1. NCBI Submission Template: BioProject and Organization
    Fig 1. NCBI Submission Template: BioProject and Organization
    ABC
    Name of Field in BioNumerics TemplateDescription  Example
    BioProject accessionIdentifier for NCBI data collection that contatins data associated to GenomeTrakr.  Specific for organism and lab submitterPRJNA514285
    Organization nameSurveillance Program (example is the default value for GenomeTrakr submissions)GenomeTrakr
    SPUID namespaceSurveillance Program (example is the default value for GenomeTrakr submissions)GenomeTrakr
    Typeorganization type (example is the default value for GenomeTrakr submissions)consortium
    Rolelaboratory responsibility (example is the default value for GenomeTrakr submissions)owner
    Contact first nameFirst name for Lab POC for NCBI submissions.  Lab might choose to create alias name for WGS teamFirst Name
    Contact last nameLast name Lab POC for NCBI submissions. Lab might choose to create alias name for WGS teamLast Name
    Contact e-mailemail for Lab POC for NCBI submissions. Lab might choose to create alias name for WGS teamfirst.last@lab.gov
    FTP upload directoryName of directory at NCBI FTP site (example is the default value for GenomeTrakr submissions)submit/Production

    Table 1. Guidelines for Bioproject and Organization metadata

    2.2.

    Laboratories will be submitting to specific bioprojects for lab/organisms. Find the organism/lab specific bioproject under each of the GenomeTrakr umbrella bioprojects included at http://www.ncbi.nlm.nih.gov/bioproject/593772

    Make sure to submit to your lab bioproject. Please don't submit to umbrella bioprojects.

    2.3.

    BioSample: Metadata associate to the isolate might require the creation of new fields in BioNumerics. The term 'field content ' field content ' denotes that the template value e.g. Organism name is mapping to the field in BioNumerics e.g. OrganismName. The template values might map to default values e.g. Pathogen: environmental/food/other; version 1.0. Make sure to include the metadata associated to the isolates in the mandatory fields such as: Submitter Provided Unique ID, BioSample accession (output), Organism name, Title, Attribute package, Strain name, Isolate name alias and Project name. Isolate name alias is a mandatory field for GenomeTrakr submissions . Provide serovar when available.

    Fig 2. NCBI Submission Template:BioSample
    Fig 2. NCBI Submission Template:BioSample
    ABCD
    Name of Field in BioNumerics TemplateDescription  Name of Field in BioNumerics DataBaseExample of metadata value
    Submitter Provided Unique ID Local lab strain ID  Entry Key21B00181-5
    BioSample accession (output)NCBI accession will get populated upon submission to NCBINCBI_ACCESSION (field content)     SAMN17385051 
    Organism nameGenus – species for organism  Organism_name (field content)Listeria monocytogenes     
    TitleOrganism nameOrganism_name (field content)Listeria monocytogenes
    Attribute packageSample category  Pathogen: environmental/food/other; version 1.0Pathogen: environmental/food/other; version 1.0
    Strain namePNUSA identifier (automatically populates at the time of registration)WGS_id (field content)PNUSAL008933
    Serovar (optional)Serotyping information for Escherichia coli and Salmonella entericaSerovar (field content)missing
    Isolate (optional)Field is not required for GenomeTrakrmissing
    Isolate namea alias (optional)Optional identifier for collaboration projectsIsolate_name_alias (field content)21B00181-5; RS_21290
    Project nameName of the project within which the sequencing was organizedProjectName (field content)

    Table 2. Guidelines for BioSample metadata

    2.4.

    BioSample: Make sure to include the metadata associated to the isolates in the mandatory fields such as: Collected by, Collection / Isolate date, Collection / Isolate date format, Title, Geographical origin, Isolate source, Sequenced by and Source name/type. Isolate name alias is a mandatory field for GenomeTrakr submissions. Provide Geographical coordinates when available. Host or host disease are provided only for isolates obtained from human, indicate "missing" for isolates from food or environmental sources.

    Fig 3. NCBI Submission Template: BioSample 2
    Fig 3. NCBI Submission Template: BioSample 2
    ABCD
    Name of Field in BioNumerics TemplateDescription  Name of Field in BioNumerics DataBaseExample of metadata value
    Collected byFull name of laboratory that collected the sample or has taken over curation of the isolate.NCBI_LabID (field content)  NY Department of Agriculture and Markets
    Collection dateDate on which the sample was collected.IsolateDate (field content)     2020
    Geographical Country and State for sample collection  SourceCountryState (field  content)     USA:NY
    Geographical coordinateslatitude and longitude for site of collection.  Missing if it is not providedmissing
    Isolation sourceDetailed description for sample product or environmental source SourceSite (field content)     cheese
    HostOnly provided for human isolates   missing
    Host diseaseOnly provided for human isolates  missing
    Sequenced byThe name of the agency that generated the sequence, e.g., Centers for Disease Control and PreventionSequenced by (field content)
    Source name / typeControlled vocabulary describing the isolation_source. Choose the best fit term: Human, Animal, Food, Environmental, Other.SourceType (field content)

    Table 3. Guidelines for BioSample metadata (2)

    2.5.

    NCBI submission settings – SRA Experiment and Run

    Populate fields for SRA Experiment and Run according to PulseNet instructions.

    Fig 4. NCBI Submission Template forBioNumerics, SRA Experiment and run:  Make sure to map collection attributes to the corresponding fields.
    Fig 4. NCBI Submission Template forBioNumerics, SRA Experiment and run: Make sure to map collection attributes to the corresponding fields.
    3.

    NCBI submission settings – Submission Template

    Save submission template according to PulseNet Instructions as - GenomeTrakr-Template -.

    4.

    Import data

    4.1.

    Import the GenomeTrakr Metadata form for BioNumerics ( GT_BioNumerics_spreadsheet_v2 .csv ) according to PulseNet Instructions.

    4.2.

    When importing rules, the field source should match destination fields.

    4.3.

    In the importing links section, choose the -key- for linking records to database entries.

    4.4.

    Proceed with sequencing data import according to PulseNet Instructions.

    4.5.

    Submit data to NCBI according to PulseNet Instructions. If NCBI accessions are not available at BioNumerics in 1 business day, please contact NCBI and PulseNet to troubleshoot issues with submissions.

    4.6.

    Contact GenomeTrakr by email genometrakr@fda.hhs.gov if issues with submissions are delayed for more than 3 days. GenomeTrakr can support urgent submissions if needed.

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