The primary workflow for the Earth Biogenome Project Pilot at NBIS.

General aim:

flowchart LR
    hifi[/ HiFi reads /] --> data_inspection
    ont[/ ONT reads /] -->  data_inspection
    hic[/ Hi-C reads /] --> data_inspection
    data_inspection[[ Data inspection ]] --> preprocessing
    preprocessing[[ Preprocessing ]] --> assemble
    assemble[[ Assemble ]] --> validation
    validation[[ Assembly validation ]] --> curation
    curation[[ Assembly curation ]] --> validation

Current implementation:

flowchart TD
    hifi[/ HiFi reads /] --> fastk_hifi[[ FastK - HiFi ]]
    hic[/ Hi-C reads /] --> fastk_hic[[ FastK - HiC ]]
    assembly[/ Assembly /] --> quast[[ Quast ]]
    fastk_hifi --> histex[[ Histex ]]
    histex --> genescopefk[[ GeneScopeFK ]]
    fastk_hifi --> ploidyplot[[ PloidyPlot ]]
    fastk_hifi --> katgc[[ KatGC ]]
    fastk_hifi --> merquryfk[[ MerquryFK ]]
    assembly --> merquryfk
    fastk_hifi --> katcomp[[ KatComp ]]
    fastk_hic --> katcomp
    assembly --> busco[[ Busco ]]
    assembly --> inspector[[ Inspector ]]
    hifi --> inspector
    refseq_sketch[( RefSeq sketch )] --> mash_screen[[ Mash Screen ]]
    hifi --> mash_screen

nextflow run -params-file <params.yml> \
    [ -c <custom.config> ] \
    [ -profile <profile> ] \
    NBISweden/Earth-Biogenome-Project-pilot

where:

• params.yml is a YAML formatted file containing workflow parameters such as input paths to the data. A params.yml template is provided to copy for convenience. Alternatively parameters can be provided on the command-line using the --parameter notation (e.g., --input <path> ).
• <custom.config> is a nextflow configuration file which provides additional configuration (see the custom.config template).
• <profile> is one of the preconfigured execution profiles (uppmax, singularity_local, docker_local). Alternatively, you can provide a custom configuration to configure this workflow to your execution environment. See Nextflow Configuration for more details.

Tool specific (module) parameters are supplied in the modules file. These can be overridden in the nextflow.config in the analysis directory, by using process selectors. For example:

// Module settings
process {
    withName: 'FASTQC' {
        // Override ext.args to FastQC
        ext.args = '--quiet'
    }
}

Mandatory:

• input: A YAML formatted input file.

Optional:

• outdir: The publishing path for results (default: results).

• publish_mode: (values: 'symlink' (default), 'copy') The file publishing method from the intermediate results folders (see Table of publish modes).

• kmer_size: The size of k-mer to use for histogram plotting.

• ploidy: The estimated ploidy.

• steps: The workflow steps to execute ( default is all steps. Choose from [data_qc,preprocess,assemble,validate,curate] ).

  Software specific:

  • multiqc_config: Path to MultiQC configuration file (default: configs/multiqc_conf.yaml).

  Software package manager specific:

  • enable_conda: Set to true to use conda as the software package manager (default: false).
  • singularity_pull_docker_container: Set to true if Singularity images should be built from the docker images, instead of retrieving existing Singularity images (default: false).

  Uppmax cluster specific:

  • project: SNIC Compute allocation number.

All results are published to the path assigned to the workflow parameter results.

• 01_FastQC_sequence_check/: FastQC output
• 02_Fastp_trimmed_sequences/: FastP output
• pipeline_info/: A folder containing workflow execution details.
• multiqc_report.html: A MultiQC report summarising the results of data processing.

A custom profile named uppmax is available to run this workflow specifically on UPPMAX clusters. The process executor is slurm so jobs are submitted to the Slurm Queue Manager. All jobs submitted to slurm must have a project allocation. This is automatically added to the clusterOptions in the uppmax profile. All Uppmax clusters have node local disk space to do computations, and prevent heavy input/output over the network (which slows down the cluster for all). The path to this disk space is provided by the $SNIC_TMP variable, used by the process.scratch directive in the uppmax profile. Lastly the profile enables the use of Singularity so that all processes must be executed within Singularity containers. See nextflow.config for the profile specification.

The profile is enabled using the -profile parameter to nextflow:

nextflow run -profile uppmax <nextflow_script>

The workflows in this folder manage the execution of your analyses from beginning to end.

workflow/
 | - bin/                            Custom workflow scripts
 | - configs/                        Configuration files that govern workflow execution
 | - containers/                     Custom container definition files
 | - docs/                           Workflow usage and interpretation information
 | - modules/                        Process definitions for tools used in the workflow
 | - subworkflows/                   Custom workflows for different stages of the main analysis
 | - main.nf                         The primary analysis script
 | - nextflow.config                 General Nextflow configuration
 \ - params.config.TEMPLATE          A Template for parameter configuration