nf-core/mag
Assembly and binning of metagenomes
2.1.0
). The latest
stable release is
3.2.1
.
Define where the pipeline should find input data and save output data.
Input FastQ files or CSV samplesheet file containing information about the samples in the experiment.
string
Use this to specify the location of your input FastQ files. For example:
--input 'path/to/data/sample_*_{1,2}.fastq.gz'
Alternatively, to assign different groups or to include long reads for hybrid assembly with metaSPAdes, you can specify a CSV samplesheet input file with 5 columns and the following header: sample,group,short_reads_1,short_reads_2,long_reads. See usage docs.
Specifies that the input is single-end reads.
boolean
By default, the pipeline expects paired-end data. If you have single-end data, you need to specify --single_end
on the command line when you launch the pipeline. A normal glob pattern, enclosed in quotation marks, can then be used for --input
. For example:
--single_end --input '*.fastq'
It is not possible to run a mixture of single-end and paired-end files in one run.
Path to the output directory where the results will be saved.
string
./results
Email address for completion summary.
string
^([a-zA-Z0-9_\-\.]+)@([a-zA-Z0-9_\-\.]+)\.([a-zA-Z]{2,5})$
Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (~/.nextflow/config
) then you don't need to specify this on the command line for every run.
MultiQC report title. Printed as page header, used for filename if not otherwise specified.
string
Reference genome related files and options required for the workflow.
Directory / URL base for iGenomes references.
string
s3://ngi-igenomes/igenomes
Do not load the iGenomes reference config.
boolean
Do not load igenomes.config
when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in igenomes.config
.
Parameters used to describe centralised config profiles. These should not be edited.
Git commit id for Institutional configs.
string
master
Base directory for Institutional configs.
string
https://raw.githubusercontent.com/nf-core/configs/master
If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.
Institutional configs hostname.
string
Institutional config name.
string
Institutional config description.
string
Institutional config contact information.
string
Institutional config URL link.
string
Set the top limit for requested resources for any single job.
Maximum number of CPUs that can be requested for any single job.
integer
16
Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. --max_cpus 1
Maximum amount of memory that can be requested for any single job.
string
128.GB
^\d+(\.\d+)?\.?\s*(K|M|G|T)?B$
Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. --max_memory '8.GB'
Maximum amount of time that can be requested for any single job.
string
240.h
^(\d+\.?\s*(s|m|h|day)\s*)+$
Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. --max_time '2.h'
Less common options for the pipeline, typically set in a config file.
Display help text.
boolean
Method used to save pipeline results to output directory.
string
The Nextflow publishDir
option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See Nextflow docs for details.
Email address for completion summary, only when pipeline fails.
string
^([a-zA-Z0-9_\-\.]+)@([a-zA-Z0-9_\-\.]+)\.([a-zA-Z]{2,5})$
An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.
Send plain-text email instead of HTML.
boolean
File size limit when attaching MultiQC reports to summary emails.
string
25.MB
^\d+(\.\d+)?\.?\s*(K|M|G|T)?B$
Do not use coloured log outputs.
boolean
Custom config file to supply to MultiQC.
string
Directory to keep pipeline Nextflow logs and reports.
string
${params.outdir}/pipeline_info
Boolean whether to validate parameters against the schema at runtime
boolean
true
Show all params when using --help
boolean
Run this workflow with Conda. You can also use '-profile conda' instead of providing this parameter.
boolean
Instead of directly downloading Singularity images for use with Singularity, force the workflow to pull and convert Docker containers instead.
boolean
This may be useful for example if you are unable to directly pull Singularity containers to run the pipeline due to http/https proxy issues.
Use these parameters to also enable reproducible results from the individual assembly and binning tools .
Fix number of CPUs for MEGAHIT to 1. Not increased with retries.
boolean
MEGAHIT only generates reproducible results when run single-threaded.
When using this parameter do not change the number of CPUs for the megahit
process with a custom config file. This would result in an error.
Default: The number of CPUs is specified in the base.config
file, and increased with each retry.
Fix number of CPUs used by SPAdes. Not increased with retries.
integer
-1
SPAdes is designed to be deterministic for a given number of threads. To generate reproducible results fix the number of CPUs using this parameter.
When using this parameter do not change the number of CPUs for the spades
process with a custom config file. This would result in an error.
Default: -1 (the number of CPUs is specified in the base.config
or in a custom config file, and increased with each retry).
Fix number of CPUs used by SPAdes hybrid. Not increased with retries.
integer
-1
SPAdes is designed to be deterministic for a given number of threads. To generate reproducible results fix the number of CPUs using this parameter.
When using this parameter do not change the number of CPUs for the spadeshybrid
process with a custom config file. This would result in an error.
Default: -1 (the number of CPUs is specified in the base.config
or in a custom config file, and increased with each retry).
RNG seed for MetaBAT2.
integer
1
MetaBAT2 is run by default with a fixed seed within this pipeline, thus producing reproducible results. You can set it also to any other positive integer to ensure reproducibility. Set the parameter to 0 to use a random seed.
Save the by fastp trimmed FastQ files in the results directory.
boolean
By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete.
Minimum phred quality value of a base to be qualified.
integer
15
The mean quality requirement used for per read sliding window cutting by fastp.
integer
15
Name of iGenomes reference for host contamination removal.
string
This parameter is mutually exclusive with --host_genome
. Host read removal is done with Bowtie2.
Both the iGenomes FASTA file as well as corresponding, already pre-built Bowtie 2 index files will be used.
Fasta reference file for host contamination removal.
string
This parameter is mutually exclusive with --host_fasta
. The reference can be masked. Host read removal is done with Bowtie2.
Use the --very-sensitive
instead of the--sensitive
setting for Bowtie 2 to map reads against the host genome.
boolean
Save the read IDs of removed host reads.
boolean
Keep reads similar to the Illumina internal standard PhiX genome.
boolean
Genome reference used to remove Illumina PhiX contaminant reads.
string
${baseDir}/assets/data/GCA_002596845.1_ASM259684v1_genomic.fna.gz
Skip removing adapter sequences from long reads.
boolean
Discard any read which is shorter than this value.
integer
1000
Keep this percent of bases.
integer
90
The higher the more important is read length when choosing the best reads.
integer
10
The default value focuses on length instead of quality to improve assembly size.
In order to assign equal weights to read lengths and read qualities set this parameter to 1.
This might be useful, for example, to benefit indirectly from the removal of short host reads (causing lower qualities for reads not overlapping filtered short reads).
Keep reads similar to the ONT internal standard Escherichia virus Lambda genome.
boolean
Genome reference used to remove ONT Lambda contaminant reads.
string
${baseDir}/assets/data/GCA_000840245.1_ViralProj14204_genomic.fna.gz
Taxonomic classification is disabled by default. You have to specify one of the options below to activate it.
Database for taxonomic binning with centrifuge.
string
E.g. ftp://ftp.ccb.jhu.edu/pub/infphilo/centrifuge/data/p_compressed+h+v.tar.gz.
Database for taxonomic binning with kraken2.
string
The database file must be a compressed tar archive that contains at least the three files hash.k2d
, opts.k2d
and taxo.k2d
. E.g. ftp://ftp.ccb.jhu.edu/pub/data/kraken2_dbs/minikraken_8GB_202003.tgz.
Skip creating a krona plot for taxonomic binning.
boolean
Database for taxonomic classification of metagenome assembled genomes.
string
E.g. https://tbb.bio.uu.nl/bastiaan/CAT_prepare/CAT_prepare_20210107.tar.gz. This parameter is mutually exclusive with --cat_db_generate
. The zipped file needs to contain a folder named *taxonomy*
and *database*
that hold the respective files.
Generate CAT database.
boolean
Download the taxonomy files from NCBI taxonomy, the nr database and generate CAT database. This parameter is mutually exclusive with --cat_db
. Useful to build a CAT database with the same DIAMOND version as used for running CAT classification, avoiding compatibility problems.
Save the CAT database generated when specified by --cat_db_generate
.
boolean
Useful to allow reproducibility, as old versions of prebuild CAT databases do not always remain accessible and underlying NCBI taxonomy and nr databases change.
GTDB database for taxonomic classification of bins with GTDB-tk.
string
https://data.gtdb.ecogenomic.org/releases/release202/202.0/auxillary_files/gtdbtk_r202_data.tar.gz
For information which GTDB reference databases are compatible with the used GTDB-tk version see https://ecogenomics.github.io/GTDBTk/installing/index.html#gtdb-tk-reference-data.
Min. bin completeness (in %) required to apply GTDB-tk classification.
number
50
Completeness assessed with BUSCO analysis (100% - %Missing). Must be greater than 0 (min. 0.01) to avoid GTDB-tk errors. If too low, GTDB-tk classification results can be impaired due to not enough marker genes!
Max. bin contamination (in %) allowed to apply GTDB-tk classification.
number
10
Contamination approximated based on BUSCO analysis (%Complete and duplicated). If too high, GTDB-tk classification results can be impaired due to contamination!
Min. fraction of AA (in %) in the MSA for bins to be kept.
number
10
Min. alignment fraction to consider closest genome.
number
0.65
Number of CPUs used for the by GTDB-Tk run tool pplacer.
number
1
A low number of CPUs helps to reduce the memory required/reported by GTDB-Tk. See also the GTDB-Tk documentation.
Reduce GTDB-Tk memory consumption by running pplacer in a setting writing to disk.
boolean
true
Will be slower. Set to false
to turn this off.
Co-assemble samples within one group, instead of assembling each sample separately.
boolean
Additional custom options for SPAdes.
string
An example is adjusting k-mers ("-k 21,33,55,77") or adding advanced options. But not -t, -m, -o or --out-prefix, because these are already in use.
Additional custom options for MEGAHIT.
string
An example is adjusting presets (e.g. "--presets meta-large"), k-mers (e.g. "-k 21,33,55,77") or adding other advanced options. For example, increase the minimum k-mer in the event of an error message such as "Too many vertices in the unitig graph, you may increase the kmer size to remove tons of erroneous kmers." in the MEGAHIT log file. But not --threads, --memory, -o or input read files, because these are already in use.
Skip Illumina-only SPAdes assembly.
boolean
Skip SPAdes hybrid assembly.
boolean
Skip MEGAHIT assembly.
boolean
Skip metaQUAST.
boolean
Defines mapping strategy to compute co-abundances for binning, i.e. which samples will be mapped against the assembly.
string
group
Available: all
, group
or own
. Note that own
cannot be specified in combination with --coassemble_group
.
Note that specifying all
without additionally specifying --coassemble_group
results in n^2
mapping processes for each assembly method, where n
is the number of samples.
Skip metagenome binning.
boolean
Minimum contig size to be considered for binning and for bin quality check.
integer
1500
For forwarding into downstream analysis, i.e. QUAST and BUSCO, and reporting.
Minimal length of contigs that are not part of any bin but treated as individual genome.
integer
1000000
Contigs that do not fulfill the thresholds of --min_length_unbinned_contigs
and --max_unbinned_contigs
are pooled for downstream analysis and reporting, except contigs that also do not fullfill --min_contig_size
are not considered further.
Maximal number of contigs that are not part of any bin but treated as individual genome.
integer
100
Contigs that do not fulfill the thresholds of --min_length_unbinned_contigs
and --max_unbinned_contigs
are pooled for downstream analysis and reporting, except contigs that also do not fullfill --min_contig_size
are not considered further.
Disable bin QC with BUSCO.
boolean
Download path for BUSCO lineage dataset, instead of using automated lineage selection.
string
E.g. https://busco-data.ezlab.org/v5/data/lineages/bacteria_odb10.2020-03-06.tar.gz. Available databases are listed here: https://busco-data.ezlab.org/v5/data/lineages/.
Path to local folder containing already downloaded and unpacked lineage datasets.
string
If provided, BUSCO analysis will be run in offline mode. Data can be downloaded from https://busco-data.ezlab.org/v5/data/ (files still need to be unpacked manually). Run in combination with automated lineage selection.
Run BUSCO with automated lineage selection, but ignoring eukaryotes (saves runtime).
boolean
Save the used BUSCO lineage datasets provided via --busco_reference or downloaded when not using --busco_reference or --busco_download_path.
boolean
Useful to allow reproducibility, as BUSCO datasets are frequently updated and old versions do not always remain accessible.