luokai0 ENA Database
Overview
The European Nucleotide Archive (ENA) is a comprehensive public repository for nucleotide sequence data and associated metadata. Access and query DNA/RNA sequences, raw reads, genome assemblies, and functional annotations through REST APIs and FTP for genomics and bioinformatics pipelines.
When to Use This Skill
This skill should be used when:
- Retrieving nucleotide sequences or raw sequencing reads by accession
- Searching for samples, studies, or assemblies by metadata criteria
- Downloading FASTQ files or genome assemblies for analysis
- Querying taxonomic information for organisms
- Accessing sequence annotations and functional data
- Integrating ENA data into bioinformatics pipelines
- Performing cross-reference searches to related databases
- Bulk downloading datasets via FTP or Aspera
Core Capabilities
1. Data Types and Structure
ENA organizes data into hierarchical object types:
Studies/Projects - Group related data and control release dates. Studies are the primary unit for citing archived data.
Samples - Represent units of biomaterial from which sequencing libraries were produced. Samples must be registered before submitting most data types.
Raw Reads - Consist of:
- Experiments: Metadata about sequencing methods, library preparation, and instrument details
- Runs: References to data files containing raw sequencing reads from a single sequencing run
Assemblies - Genome, transcriptome, metagenome, or metatranscriptome assemblies at various completion levels.
Sequences - Assembled and annotated sequences stored in the EMBL Nucleotide Sequence Database, including coding/non-coding regions and functional annotations.
Analyses - Results from computational analyses of sequence data.
Taxonomy Records - Taxonomic information including lineage and rank.
2. Programmatic Access
ENA provides multiple REST APIs for data access. Consult references/api_reference.md for detailed endpoint documentation.
Key APIs:
ENA Portal API - Advanced search functionality across all ENA data types
- Documentation: https://www.ebi.ac.uk/ena/portal/api/doc
- Use for complex queries and metadata searches
ENA Browser API - Direct retrieval of records and metadata
- Documentation: https://www.ebi.ac.uk/ena/browser/api/doc
- Use for downloading specific records by accession
- Returns data in XML format
ENA Taxonomy REST API - Query taxonomic information
- Access lineage, rank, and related taxonomic data
ENA Cross Reference Service - Access related records from external databases
- Endpoint: https://www.ebi.ac.uk/ena/xref/rest/
CRAM Reference Registry - Retrieve reference sequences
- Endpoint: https://www.ebi.ac.uk/ena/cram/
- Query by MD5 or SHA1 checksums
Rate Limiting: All APIs have a rate limit of 50 requests per second. Exceeding this returns HTTP 429 (Too Many Requests).
3. Searching and Retrieving Data
Browser-Based Search:
- Free text search across all fields
- Sequence similarity search (BLAST integration)
- Cross-reference search to find related records
- Advanced search with Rulespace query builder
Programmatic Queries:
- Use Portal API for advanced searches at scale
- Filter by data type, date range, taxonomy, or metadata fields
- Download results as tabulated metadata summaries or XML records
Example API Query Pattern:
import requests
# Search for samples from a specific study
base_url = "https://www.ebi.ac.uk/ena/portal/api/search"
params = {
"result": "sample",
"query": "study_accession=PRJEB1234",
"format": "json",
"limit": 100
}
response = requests.get(base_url, params=params)
samples = response.json()
4. Data Retrieval Formats
Metadata Formats:
- XML (native ENA format)
- JSON (via Portal API)
- TSV/CSV (tabulated summaries)
Sequence Data:
- FASTQ (raw reads)
- BAM/CRAM (aligned reads)
- FASTA (assembled sequences)
- EMBL flat file format (annotated sequences)
Download Methods:
- Direct API download (small files)
- FTP for bulk data transfer
- Aspera for high-speed transfer of large datasets
- enaBrowserTools command-line utility for bulk downloads
5. Common Use Cases
Retrieve raw sequencing reads by accession:
# Download run files using Browser API
accession = "ERR123456"
url = f"https://www.ebi.ac.uk/ena/browser/api/xml/{accession}"
Search for all samples in a study:
# Use Portal API to list samples
study_id = "PRJNA123456"
url = f"https://www.ebi.ac.uk/ena/portal/api/search?result=sample&query=study_accession={study_id}&format=tsv"
Find assemblies for a specific organism:
# Search assemblies by taxonomy
organism = "Escherichia coli"
url = f"https://www.ebi.ac.uk/ena/portal/api/search?result=assembly&query=tax_tree({organism})&format=json"
Get taxonomic lineage:
# Query taxonomy API
taxon_id = "562" # E. coli
url = f"https://www.ebi.ac.uk/ena/taxonomy/rest/tax-id/{taxon_id}"
6. Integration with Analysis Pipelines
Bulk Download Pattern:
- Search for accessions matching criteria using Portal API
- Extract file URLs from search results
- Download files via FTP or using enaBrowserTools
- Process downloaded data in pipeline
BLAST Integration: Integrate with EBI's NCBI BLAST service (REST/SOAP API) for sequence similarity searches against ENA sequences.
7. Best Practices
Rate Limiting:
- Implement exponential backoff when receiving HTTP 429 responses
- Batch requests when possible to stay within 50 req/sec limit
- Use bulk download tools for large datasets instead of iterating API calls
Data Citation:
- Always cite using Study/Project accessions when publishing
- Include accession numbers for specific samples, runs, or assemblies used
API Response Handling:
- Check HTTP status codes before processing responses
- Parse XML responses using proper XML libraries (not regex)
- Handle pagination for large result sets
Performance:
- Use FTP/Aspera for downloading large files (>100MB)
- Prefer TSV/JSON formats over XML when only metadata is needed
- Cache taxonomy lookups locally when processing many records
Resources
This skill includes detailed reference documentation for working with ENA:
references/
api_reference.md - Comprehensive API endpoint documentation including:
- Detailed parameters for Portal API and Browser API
- Response format specifications
- Advanced query syntax and operators
- Field names for filtering and searching
- Common API patterns and examples
Load this reference when constructing complex API queries, debugging API responses, or needing specific parameter details.