Skip to content

Latest commit

 

History

History
127 lines (103 loc) · 40.8 KB

Genome_Technical_Metadata.md

File metadata and controls

127 lines (103 loc) · 40.8 KB

2.1 Genome sequencing

A searchable and exportable tab-separated table of the following metadata is now available.

Minimal technical metadata for Genomic FASTQ data

metadata definition reference of definition[<url_to_definition>] expected unit of measurement example sources (where this or similar matadata field is mentioned)
samp_name A local identifier or name that for the material sample used for extracting nucleic acids, and subsequent sequencing. It can refer either to the original material collected or to any derived sub-samples. It can have any format, but we suggest that you make it concise, unique and consistent within your lab, and as informative as possible. INSDC requires every sample name from a single Submitter to be unique. MIXS:0001107 free text with identifier e.g. ISDsoil1 GSC MIxS/MIGS Bacteria (“GSC MIXS: MIGSBacteria”)
seq_meth Sequencing machine used. Where possible the term should be taken from the OBI list of DNA sequencers (http://purl.obolibrary.org/obo/OBI_0400103) MIXS:0000050 <name_of_seq_machine>[ontology] e.g. 454 Genome Sequencer FLX [OBI:0000702] GSC MIxS/MIGS Bacteria (“GSC MIXS: MIGSBacteria”), ENA Metadata Validation: Instrument (“ENA Metadata Validation: Instrument”)
lib_layout Specify whether to expect single, paired, or other configuration of reads MIXS:0000041 free text string e.g. single-end GSC MIxS/MIGS Bacteria (“GSC MIXS: MIGSBacteria”)
lib_source The lib_source specifies the type of source material that is being sequenced Link to permitted values Free text from selected list of values e.g. GENOMIC, METAGENOMIC, TRANSCRIPTOMIC, etc. ENA Metadata Validation: Source (“ENA Metadata Validation: Source”)
lib_strategy Sequencing technique intended for this library Link to permitted values Free text from selected list of values e.g. WGS, WGA, Amplicon, etc. ENA Metadata Validation: Strategy (“ENA Metadata Validation: Strategy”)
lib_selection Whether any method was used to select and/or enrich the material being sequenced Link to permitted values Free text from selected list of values e.g. RANDOM, PCR, cDNA_oligo_dT etc. ENA Metadata Validation: Selection (“ENA Metadata Validation: Selection”)
spots Number of reads in the library (sequencing depth), assigned at submission Link to submission of genomes integer value + unit of measurement e.g. 32,283,453 OR 32.3M Adapted from NCBI-SRA (Leinonen et al. 2011)
bases Number of base pairs (nucleotides) in the library, assigned at submission Link to submission of genomes integer value + unit of measurement e.g. 6,400,000 or 6.4M Adapted from NCBI-SRA (Leinonen et al. 2011)
nucl_acid_ext A link to a literature reference, electronic resource or a standard operating procedure (SOP), that describes the material separation to recover the nucleic acid fraction from a sample MIXS:0000037 PMID, DOI, URL e.g. https://doi.org/10.1038/s41586-020-2192-1 GSC MISAG [gsc_migs_bacteria]
nucl_acid_amp A link to a literature reference, electronic resource or a standard operating procedure (SOP), that describes the enzymatic amplification (PCR, TMA, NASBA) of specific nucleic acids MIXS:0000038 PMID, DOI, URL e.g. https://phylogenomics.me/protocols/16s-pcr-protocol/ GSC MISAG [gsc_migs_bacteria]
average_length As bases divided by spots, calculated by user Link to use in databse Integer value calculated by tool e.g. 198 Calculated as basepairs_count/sequence_count
sequence_count_qual (optional) Number of reads in the library (sequencing depth) after quality filtering Link to resource to calculate integer value + % SRA-Tinder (NCBI Hackathons)
basepairs_count_qual (optional) Number of base pairs (nucleotides) in the library after quality filtering Link to resource to calculate integer value + % SRA-Tinder (NCBI Hackathons)
checksum Hash value for data integrity Link to ENA MD5 Checksum string with checksum e.g. MD5: cbc41d0e49636872a765b950cb7f410a Data transfer and data integrity

Minimal technical metadata for Genomic FASTA data

metadata definition reference of definition[<url_to_definition>] expected unit of measurement example sources( where this or similar matadata field is mentioned)
run_ref Accessions/identifiers linking to the raw data (FASTQ) Link to reference run_accession in the format SRR, ERR or DRR e.g. RUN_REF accession = “ERR178314” Adapted from ENA (“ENA How to Submit Other Analyses: Submitting Read Alignments”)
tax_ident The phylogenetic marker(s) used to assign an organism name to the genome MIXS:0000053 free text e.g. 16s rRNA gene, multi-marker approach, other GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)
assembly_qual The assembly quality category is based on sets of criteria outlined for each assembly quality category. For MISAG/MIMAG; Finished: Single, validated, contiguous sequence per replicon without gaps or ambiguities with a consensus error rate equivalent to Q50 or better. High Quality Draft:Multiple fragments where gaps span repetitive regions. Presence of the large subunit (LSU) RNA, small subunit (SSU) and the presence of 5.8S rRNA or 5S rRNA depending on whether it is a eukaryotic or prokaryotic genome, respectively. Medium Quality Draft:Many fragments with little to no review of assembly other than reporting of standard assembly statistics. Low Quality Draft:Many fragments with little to no review of assembly other than reporting of standard assembly statistics. Assembly statistics include, but are not limited to total assembly size, number of contigs, contig N50/L50, and maximum contig length. Genome fragment(s): One or multiple fragments, totalling < 90% of the expected genome or replicon sequence, or for which no genome size could be estimated MIXS:0000056 free text from predetermined strings e.g. Medium Quality Draft GSC MIxS/MIGS Bacteria (“GSC MIXS: MIGSBacteria”)
assembly_software Tool(s) used for assembly, including version number and parameters MIXS:0000058 free text e.g. metaSPAdes (3.11.0);kmer set 21,33,55,77,99,121, default parameters otherwise GSC MIxS/MIGS Bacteria (“GSC MIXS: MIGSBacteria”)
coverage The estimated depth of sequencing coverage (in x) Link to usecase integer value 30.5 ENA Submitting Metagenome Assemblies (“ENA Submitting Metagenome Assemblies”)
number_contig Total number of contigs in the cleaned/submitted assembly that makes up a given genome, SAG, MAG, or UViG MIXS:0000060 integer value e.g. 40 GSC MIxS/MIGS Bacteria (“GSC MIXS: MIGSBacteria”), Roadmap for naming uncultivated Archaea and Bacteria (Murray et al. 2020)
N50 The length of the shortest contig representing half of the assembly length Link to reference 1 Link to reference 2 integer value + unit e.g. N50=4kb Roadmap for naming uncultivated Archaea and Bacteria (Murray et al. 2020)
x16s_recover Can a 16S gene be recovered from the submitted sequence MIXS:0000065 free text string e.g. yes Adapted from GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)
x16s_recover_software Tools used for 16S rRNA gene extraction MIXS:0000066 free text string e.g. rambl (v2); default parameters Adapted from GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)
trnas Total number of tRNAs identified from the genome MIXS:0000067 integer value e.g. 18 GSC MIXS: MIMAG (“GSC MIXS: MIMAG”), Roadmap for naming uncultivated Archaea and Bacteria (Murray et al. 2020)
trna_ext_software Tools used for tRNA identification MIXS:0000068 free text string e.g. infernal (v2); default parameters GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)
compl_score Completeness score is typically based on either the fraction of markers found as compared to a database or the percent of a genome found as compared to a closely related reference genome. High Quality Draft: >90%, Medium Quality Draft: >50%, and Low Quality Draft: < 50% should have the indicated completeness scores MIXS:0000069 integer value (%) e.g. med; 60% GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)
compl_software Tools used for completion estimate MIXS:0000070 free text string e.g. checkm (v1.1.6) GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)
contam_score The contamination score is based on the fraction of single-copy genes that are observed more than once in a query genome. The following scores are acceptable for; High Quality Draft: < 5%, Medium Quality Draft: < 10%, Low Quality Draft: < 10%. Contamination must be below 5% for a SAG or MAG to be deposited into any of the public databases. MIXS:0000072 integer value (%) e.g. 0.01 GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)
decontam_software Tool(s) used in contamination screening MIXS:0000074 free text string e.g. checkm (v1.1.6) GSC MIXS: MIMAG (“GSC MIXS: MIMAG”)

The publications describing the reasons for formation of The minimum information about a genome sequence (MIGS) (Field et al. 2008), Minimum information about a marker gene sequence (MIMARKS) and minimum information about any (x) sequence (MIxS) specifications (Yilmaz et al. 2011), Minimum information about a single amplified genome (MISAG) and a metagenome-assembled genome (MIMAG) of bacteria and archaea (Bowers et al. 2017), and Roadmap for naming uncultivated Archaea and Bacteria (Murray et al. 2020) can be found online.

We also encourage our readers to have a look at the HumanMetagenomeDB: a public repository of curated and standardized metadata for human metagenomes (Kasmanas et al. 2021) and TerrestrialMetagenomeDB: a public repository of curated and standardized metadata for terrestrial metagenomes (Corrêa et al. 2020) to better understand some of the metadata fields that can be found in the tables above.

References

Bowers, R., N. Kyrpides, R. Stepanauskas, et al. 2017. “Minimum Information about a Single Amplified Genome (MISAG) and a Metagenome-Assembled Genome (MIMAG) of Bacteria and Archaea.” Nat Biotechnol 35: 725–31. https://doi.org/10.1038/nbt.3893.

Corrêa, Fabio B., João P. Saraiva, Peter F. Stadler, and Ulisses N. da Rocha. 2020. “TerrestrialMetagenomeDB: A Public Repository of Curated and Standardized Metadata for Terrestrial Metagenomes.” Nucleic Acids Research 48 (D1): D626–32. https://academic.oup.com/nar/article/48/D1/D626/5625925.

“ENA How to Submit Other Analyses: Submitting Read Alignments.” https://ena-docs.readthedocs.io/en/latest/submit/analyses/read-alignments.html.

“ENA Metadata Validation: Instrument.” https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#instrument.

“ENA Metadata Validation: Selection.” https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#selection.

“ENA Metadata Validation: Source.” https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#source.

“ENA Metadata Validation: Strategy.” https://ena-docs.readthedocs.io/en/latest/submit/reads/webin-cli.html#strategy.

“ENA Submitting Metagenome Assemblies.” https://ena-docs.readthedocs.io/en/latest/submit/assembly/metagenome.html.

Field, D., G. Garrity, T. Gray, N. Morrison, J. Selengut, P. Sterk, T. Tatusova, et al. 2008. “The Minimum Information about a Genome Sequence (MIGS) Specification.” Nature Biotechnology. 2008. https://doi.org/10.1038/nbt1360.

“GSC MIXS: MIGSBacteria.” https://genomicsstandardsconsortium.github.io/mixs/0010003/.

“GSC MIXS: MIMAG.” https://genomicsstandardsconsortium.github.io/mixs/MIMAG/.

Kasmanas, João C., Alexander Bartholomäus, Fabio B. Corrêa, Tal Tal, Nico Jehmlich, Gunda Herberth, Martin von Bergen, Peter F. Stadler, Alinne de Carvalho, and Ulisses N. da Rocha. 2021. “HumanMetagenomeDB: A Public Repository of Curated and Standardized Metadata for Human Metagenomes.” Nucleic Acids Research 49 (D1): D743–50. https://academic.oup.com/nar/article/49/D1/D743/5998395.

Leinonen, R., H. Sugawara, M. Shumway, and International Nucleotide Sequence Database Collaboration. 2011. “The Sequence Read Archive.” Nucleic Acids Research 39 (Database issue): D19–21. https://doi.org/10.1093/nar/gkq1019.

Murray, A. E., J. Freudenstein, S. Gribaldo, et al. 2020. “Roadmap for Naming Uncultivated Archaea and Bacteria.” Nat Microbiol 5: 987–94. https://doi.org/10.1038/s41564-020-0733-x.

NCBI Hackathons. “SRA-Tinder: A Tool to Discover Related Sequence Read Archive (SRA) Experiments.” https://github.com/NCBI-Hackathons/SRA_Tinder.

Yilmaz, Pelin et al. 2011. “Minimum Information about a Marker Gene Sequence (MIMARKS) and Minimum Information about Any (x) Sequence (MIxS) Specifications.” Nature Biotechnology 29 (5): 415–20. https://doi.org/10.1038/nbt.1823.