ENA submission · interactive

Up the wire

A finished metagenome doesn’t become public the moment you press submit. ENA’s archive isn’t a flat record — it’s a directed graph of five object types (Study, Sample, Experiment, Run, Analysis) plus a Submission control object, and a sample is only citable once every tier above it exists and has handed an accession back down. By hand you’d register each of those yourself, as XML or as a Webin-CLI manifest. SeqDesk’s submg path collapses all of it into a single per-sample YAML — where exactly two of your answers become named fields, your whole MIxS questionnaire rides up as sample attributes, and six accessions later climb back into four database rows.

What you can submit to ENA — and what SeqDesk covers

ENA accepts far more than reads and an assembly. Scroll the menu, category by category, and watch how much of it SeqDesk actually fills.

4ENA types SeqDesk covers4 of 24 shown
Metadata4/4
Sequence reads
Assemblies & genomes
Other analyses
Different archive
SeqDesk ENA only other archive
1 Metadata4 of 4 in SeqDesk

Before any sequence goes up, ENA needs the metadata objects — the project, the sample, its checklist, and the submission wrapper. SeqDesk registers all four on its XML path.

  • Study / ProjectSeqDeskgroups the work, sets the release date · Project · PRJEB
  • Sample / BioSampleSeqDeskthe biomaterial, tied to a taxon · Sample · ERS + SAMEA
  • Sample checklist (MIxS)SeqDeskthe metadata questionnaire · Sample attributes · ERC*
  • Submission (ADD · HOLD/RELEASE)SeqDeskthe action wrapper · Submission
2 Sequence reads2 of 4 in SeqDesk

Then the reads. SeqDesk submits cleaned paired-end FASTQ through submg — but ENA also takes single-end and long reads, which that path doesn't.

  • Library + instrumentvia submghow it was sequenced · Experiment · ERX
  • Paired-end reads (cleaned)via submgR1/R2 FASTQ + MD5 · Run · ERR
  • Single-end readsENA onlyone FASTQ per run · Run · ERR
  • Long reads (Nanopore / PacBio)ENA onlysingle-molecule reads · Run · ERR
3 Assemblies & genomes3 of 7 in SeqDesk

ENA accepts many kinds of assembly. SeqDesk covers the metagenome flavours — primary, binned, MAG — and leaves isolate genomes, SAGs, transcriptomes and targeted loci to ENA's own tools.

  • Primary metagenome assemblyvia submgthe community assembly · Analysis · ERZ
  • Binned metagenomevia submgsingle-taxon bins · Analysis + derived sample
  • MAG (metagenome-assembled genome)via submga recovered genome · Analysis + derived sample
  • Isolate / individual genomeENA onlya cultured-organism genome · Analysis → GCA
  • Single-cell amplified genome (SAG)ENA onlya genome from one cell · Analysis
  • Transcriptome assembly (TSA)ENA onlyassembled transcripts · Analysis
  • Targeted loci / 16S · annotated flatfilesENA onlymarker genes, EMBL flatfiles · Analysis (sequence)
4 Other analyses0 of 6 in SeqDesk

ENA's schema lists 22 analysis types — alignments, variants, annotations, AMR, phylogenies and more. None of these are in SeqDesk today.

  • Read alignment (BAM / CRAM)ENA onlyreads against a reference · Analysis · REFERENCE_ALIGNMENT
  • Sequence variation (VCF)ENA onlyvariant calls · Analysis · SEQUENCE_VARIATION
  • Sequence / functional annotationENA onlyfeature tables · Analysis · SEQUENCE_ANNOTATION
  • Taxonomic reference setENA onlya metabarcoding database · Analysis · TAXONOMIC_REFERENCE_SET
  • AMR antibiogramENA onlysusceptibility data · BioSample structured data
  • Phylogeny · genome maps · COVID-19 · TPA …ENA only+ more derived analyses · Analysis (various)
5 Different archive0 of 3 in SeqDesk

And some data was never ENA's to take: controlled human data goes to EGA, open variants to EVA, expression matrices to ArrayExpress.

  • Controlled-access human dataother archiveidentifiable human reads/variants · EGA
  • Open cross-species variantsother archiveFAIR open VCFs · EVA
  • Processed expression / microarrayother archiveRPKM matrices, study designs · ArrayExpress / BioStudies

SeqDesk’s slice is study + sample metadata, cleaned paired-end reads, and (meta)genome assemblies with bins/MAGs. Everything marked “ENA only” is submittable to ENA but outside SeqDesk; the three sister archives (EGA, EVA, ArrayExpress) are different services. ENA-side detail from the ENA submission docs and the SRA analysis schema.

The rest of this post zooms into that supported slice — a single metagenome with cleaned paired-end reads and an assembly. Even for the data SeqDesk does cover, ENA still wants the full object graph below.

The submission you would build by hand

A valid ENA submission is not a form — it is a directed graph of objects, each blocked on an accession the archive only mints once the tier above it exists. Scroll to build it; drag a node or zoom to explore.

citable tracking only control objectmetadata XML vs Webin-CLI
objects to register by hand
0 / 6
5 ENA objects + 1 Submission control

The leading letter of each accession (E, D or S) marks the INSDC partner that took the record — E for ENA/EMBL-EBI, D for DDBJ, S for NCBI. Click any node for its exact format and the SeqDesk column it writes back to.

One YAML instead of six registrations

SeqDesk doesn’t ask you to assemble that graph yourself. Its XML registration path creates the Study and the Sample; then the submg pipeline — admin-triggered, one run per study or per chosen subset of samples — writes a single YAML document for each sample and hands it to submg submit. Colour that document by who actually supplies each line and the division of labour becomes hard to miss.

Dissect the YAML — who supplies each line

The real per-sample document SeqDesk hands to submg, coloured by who fills each line. Scroll to pour the colour down; the side ledger counts each class as it appears.

your MIxS questionnaireSeqDesk scaffoldingminted by ENA, written back
STUDY: "PRJEB71234"
PROJECT_NAME: "Mouse gut recovery time-series"
SEQUENCING_PLATFORMS: ["ILLUMINA"]
METAGENOME_TAXID: "410661"
METAGENOME_SCIENTIFIC_NAME: "mouse gut metagenome"
NEW_SAMPLES:
- TITLE: "Gut recovery — day 0"
collection date: "2024-03-12"
geographic location (country and/or sea): "Germany"
ADDITIONAL_SAMPLESHEET_FIELDS:
"host scientific name": "Mus musculus"
"broad-scale environmental context": "rodent gut [ENVO]"
… and every other answer — N fields per checklist
PAIRED_END_READS:
- NAME: "GUT_D0"
PLATFORM: "ILLUMINA"
LIBRARY_NAME: "GUT_D0_library"
SEQUENCING_INSTRUMENT: "Illumina NovaSeq 6000"default
LIBRARY_SOURCE: "METAGENOMIC"default
LIBRARY_SELECTION: "RANDOM"default
LIBRARY_STRATEGY: "WGS"default
INSERT_SIZE: "300"default
FASTQ1_FILE: "…/GUT_D0_R1.fastq.gz"
FASTQ2_FILE: "…/GUT_D0_R2.fastq.gz"
RELATED_SAMPLE_TITLE: "Gut recovery — day 0"
ADDITIONAL_MANIFEST_FIELDS:
ASSEMBLY:
ASSEMBLY_NAME: "GUT_D0_assembly"
ASSEMBLY_SOFTWARE: "MEGAHIT"default
COVERAGE_VALUE: "1"default
ISOLATION_SOURCE: "UNKNOWN"default
FASTA_FILE: "…/GUT_D0.fasta"
collection date: "2024-03-12"
geographic location (country and/or sea): "Germany"
ADDITIONAL_MANIFEST_FIELDS:
BINS: # only when bins are present
BINS_DIRECTORY: "…/bins"
COMPLETENESS_SOFTWARE: "CheckM"default
QUALITY_FILE: "…/checkm_summary_GUT_D0.tsv"
BINNING_SOFTWARE: "MetaBAT2"default
ISOLATION_SOURCE: "UNKNOWN"default
MANUAL_TAXONOMY_FILE: "…/tax_ids_GUT_D0.txt"
ADDITIONAL_MANIFEST_FIELDS:
── these arrive later — written back, never authored ──
Sample.sampleAccessionNumber ← "ERS18002245"
Sample.biosampleNumber ← "SAMEA117338901"citable
Read.experimentAccessionNumber ← "ERX13002277"
Read.runAccessionNumber ← "ERR14552310"
Assembly.assemblyAccession ← "ERZ24117788"tracking
Bin.binAccession ← "ERZ24117·…"

Counts are for this illustrative sample (one read set, with bins). The ADDITIONAL_SAMPLESHEET_FIELDS block is as long as your checklist — each ENA/GSC sample checklist offers 77–125 fields — so it is shown as N per checklist, not a fixed number. Every key, default and source line is read straight from submg-runner.ts.

So the answer to does it upload the MIxS questionnaire? is: literally yes. Inside NEW_SAMPLES, two answers are hoisted into named keys — collection date and geographic location (country and/or sea) — and every other checklist answer is passed through under ADDITIONAL_SAMPLESHEET_FIELDS, which ENA stores as the sample’s SAMPLE_ATTRIBUTEs. The rest of the file is scaffolding: the platform, the taxid, the library descriptors, defaults like INSERT_SIZE 300 and ASSEMBLY_SOFTWARE MEGAHIT, resolved file paths. And the accessions — the part that makes the work citable — aren’t in the file at all. Watch them arrive.

Up the wire — accessions mint back down

As each --submit flag fires, its tier lights and ENA mints an accession back into a SeqDesk database row. Scroll to send the sample up.

Studyprerequisitemetadata XML
PRJEB71234StudyPRJ(E|D|N)[A-Z][0-9]+

Registered first by SeqDesk's XML path. submg refuses to run until the study has a PRJ accession.

Study.studyAccessionId
Sample--submit-samplesmetadata XML
ERS18002245ENA sample(E|D|S)RS[0-9]{6,}SAMEA117338901BioSample · cite thisSAM(E|D|N)[A-Z]?[0-9]+

The sample lands two accessions at once. Writing the SAMEA BioSample cross-reference back is unique to the submg path — the XML path stores only the ERS.

Sample.sampleAccessionNumberSample.biosampleNumber
Experiment--submit-readsWebin-CLI
ERX13002277Experiment · tracking(E|D|S)RX[0-9]{6,}

Records the library + instrument and joins the sample to its run.

Read.experimentAccessionNumber
Run--submit-readsWebin-CLI
ERR14552310Run · tracking(E|D|S)RR[0-9]{6,}

Carries the FASTQ files up through Webin-CLI.

Read.runAccessionNumber
Analysis · assembly--submit-assemblyWebin-CLI
ERZ24117788ERZ · tracking(E|D|S)RZ[0-9]{6,}GCA_968211045.1GCA · cite thisGCA_[0-9]{9}\.[0-9]+

Webin-CLI returns an ERZ tracking id, which SeqDesk stores in assemblyAccession. ENA issues the citable GCA later, after processing — SeqDesk does not mint or store it.

Assembly.assemblyAccession
Bins / MAGs--submit-binsWebin-CLI
ERZ24117·…per-bin(E|D|S)RZ[0-9]{6,}

Optional. Each bin is its own derived sample + analysis; a sample with no bins is a warning, not a failure.

Bin.binAccession

Accession values shown are format exemplars matching ENA’s documented regex — not real archived records. The citable GCA is issued later by ENA after processing and is not minted or stored by SeqDesk; SeqDesk records only the ERZ tracking id in Assembly.assemblyAccession.

Six accessions for one sample, landing in four SeqDesk rows. Only three are for citing — the Study (PRJEB), the Sample’s BioSample (SAMEA), and the eventual assembly (GCA); ENA lists the rest as tracking accessions, not among the citable ones. But none of them get minted at all until the sample clears SeqDesk’s pre-flight.

The gates before the wire

Nothing submits until the sample clears every hard-fail gate in prepareSubmgRun. Two of them are the MIxS fields from the YAML above.

On the Test target there is one extra gate: the study's test registration must be under 24 h old, because ENA reloads its sandbox from production every day. Click any gate to see what blocks it.

The last two gates — an empty collection date or geographic location — are the same two gold fields from the YAML we dissected. The questionnaire was never paperwork; it is the boarding pass for the wire. All gate text is transcribed from submg-runner.ts.

Data & method. Computed at build time from SeqDesk’s submg pipeline (src/lib/pipelines/submg/submg-runner.ts) and ENA’s accession documentation. The YAML structure, defaults, write-back fields and pre-flight gates are transcribed from that source with line references; the accession strings shown are format exemplars matching ENA’s documented regex, not real archived records. The ADDITIONAL_SAMPLESHEET_FIELDS passthrough is variable per checklist (shown as N), and the per-class line counts are for the single illustrative sample dissected above. submg is the open-source tool by Tubbesing et al. (BioData Mining, 2025).