Swedish Earth Biogenome Project - Genome Assembly Workflow
The primary genome assembly workflow for the Earth Biogenome Project at NBIS.
Workflow overview
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
input[/ Input file/] --> hifi
input --> hic
input --> taxonkit[[ TaxonKit name2taxid/reformat ]]
taxonkit --> goat_taxon[[ GOAT taxon search ]]
goat_taxon --> busco
goat_taxon --> dtol[[ DToL lookup ]]
hifi --> samtools_fa[[ Samtools fasta ]]
samtools_fa --> fastk_hifi
samtools_fa --> mash_screen
hifi[/ HiFi reads /] --> fastk_hifi[[ FastK - HiFi ]]
hifi --> meryl_hifi[[ Meryl - HiFi ]]
hic[/ Hi-C reads /] --> fastk_hic[[ FastK - Hi-C ]]
hifi --> meryl_hic[[ Meryl - Hi-C ]]
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
meryl_hifi --> merqury[[ Merqury ]]
assembly --> merqury
fastk_hifi --> katcomp[[ KatComp ]]
fastk_hic --> katcomp
assembly --> busco[[ Busco ]]
refseq_sketch[( RefSeq sketch )] --> mash_screen[[ Mash Screen ]]
hifi --> mash_screen
fastk_hifi --> hifiasm[[ HiFiasm ]]
hifiasm --> assembly
assembly --> purgedups[[ Purgedups ]]
input --> mitoref[[ Mitohifi - Find reference ]]
assembly --> mitohifi[[ Mitohifi ]]
assembly --> fcsgx[[ FCS GX ]]
fcs_fetchdb[( FCS fetchdb )] --> fcsgx
mitoref --> mitohifi
genescopefk --> quarto[[ Quarto ]]
goat_taxon --> multiqc[[ MultiQC ]]
quarto --> multiqc
dtol --> multiqc
katgc --> multiqc
ploidyplot --> multiqc
busco --> multiqc
quast --> multiqc
Usage
nextflow run -params-file \
[ -c ] \
[ -profile ] \
NBISweden/Earth-Biogenome-Project-pilot
where:
-
params.yml
is a YAML formatted file containing workflow parameters such as input paths to the assembly specification, and settings for tools within the workflow.Example:
input: 'assembly_spec.yml' outdir: results fastk: # Optional kmer_size: 31 # default 31 genescopefk: # Optional kmer_size: 31 # default 31 hifiasm: # Optional, default = no extra options: Key (e.g. 'opts01') is used in assembly build name (e.g., 'hifiasm-raw-opts01'). opts01: "--opts A" opts02: "--opts B" busco: # Optional, default: retrieved from GOAT lineages: 'auto' # comma separated string of lineages or auto.
Alternatively parameters can be provided on the command-line using the
--parameter
notation (e.g.,--input
). -
`` is a Nextflow configuration file which provides additional configuration. This is used to customise settings other than workflow parameters, such as cpus, time, and command-line options to tools.
Example:
process { withName: 'BUSCO' { // Selects the process to apply settings. cpus = 6 // Overrides cpu settings defined in nextflow.config time = 4.d // Overrides time settings defined in nextflow.config to 4 days. Use .h for hours, .m for minutes. memory = '20GB' // Overrides memory settings defined in nextflow.config to 20 GB. // ext.args supplies command-line options to the process tool // overrides settings found in configs/modules.config ext.args = '--long' // Supplies these as command-line options to Busco } }
-
`` is one of the preconfigured execution profiles (
uppmax
, `singularity_local`, `docker_local`, etc: see nextflow.config). Alternatively, you can provide a custom configuration to configure this workflow to your execution environment. See Nextflow Configuration for more details.
Workflow parameter inputs
Mandatory:
-
input
: A YAML formatted input file. Exampleassembly_spec.yml
(See also test profile input TODO:: Update test profile):sample: # Required: Meta data name: 'Laetiporus sulphureus' # Required: Species name. Correct spelling is important to look up species information. ploidy: 2 # Optional: Estimated ploidy (default: retrieved from GOAT) genome_size: 2345 # Optional: Estimated genome size (default: retrieved from GOAT) haploid_number: 13 # Optional: Estimated haploid chromosome count (default: retrieved from GOAT) taxid: 5630 # Optional: Taxon ID (default: retrieved with Taxonkit) kingdom: Eukaryota # Optional: (default: retrived with Taxonkit) assembly: # Optional: List of assemblies to curate and validate. - assembler: hifiasm # For each entry, the assembler, stage: raw # stage of assembly, id: uuid # unique id, pri_fasta: /path/to/primary_asm.fasta # and paths to sequences are required. alt_fasta: /path/to/alternate_asm.fasta pri_gfa: /path/to/primary_asm.gfa alt_gfa: /path/to/alternate_asm.gfa - assembler: ipa stage: raw id: uuid pri_fasta: /path/to/primary_asm.fasta alt_fasta: /path/to/alternate_asm.fasta hic: # Optional: List of hi-c reads to QC and use for scaffolding - read1: '/path/to/raw/data/hic/LS_HIC_R001_1.fastq.gz' read2: '/path/to/raw/data/hic/LS_HIC_R001_2.fastq.gz' hifi: # Required: List of hifi-reads to QC and use for assembly/validation - reads: '/path/to/raw/data/hifi/LS_HIFI_R001.bam' rnaseq: # Optional: List of Rna-seq reads to use for validation - read1: '/path/to/raw/data/rnaseq/LS_RNASEQ_R001_1.fastq.gz' read2: '/path/to/raw/data/rnaseq/LS_RNASEQ_R001_2.fastq.gz' isoseq: # Optional: List of Isoseq reads to use for validation - reads: '/path/to/raw/data/isoseq/LS_ISOSEQ_R001.bam'
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). -
steps
: The workflow steps to execute (default is all steps). Choose from:inspect
: 01 - Read inspectionpreprocess
: 02 - Read preprocessingassemble
: 03 - Assemblypurge
: 04 - Duplicate purgingpolish
: 05 - Error polishingscreen
: 06 - Contamination screeningscaffold
: 07 - Scaffoldingcurate
: 08 - Rapid curationalignRNA
: 09 - Align RNAseq data
Software specific:
Tool specific settings are provided by supplying values to specific keys or supplying an array of
settings under a tool name. The input to -params-file
would look like this:
input: assembly.yml
outdir: results
fastk:
kmer_size: 31
genescopefk:
kmer_size: 31
hifiasm:
opts01: "--opts A"
opts02: "--opts B"
busco:
lineages: 'auto'
multiqc_config
: Path to MultiQC configuration file (default:configs/multiqc_conf.yaml
).
Uppmax and PDC cluster specific:
project
: NAISS Compute allocation number.
Workflow outputs
All results are published to the path assigned to the workflow parameter results
.
TODO:: List folder contents in results file
Customization for Uppmax
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
A NAISS compute allocation should also be supplied using the --project
parameter.
Customization for PDC
A custom profile named dardel
is available to run this workflow specifically
on the PDC cluster Dardel. 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 dardel
profile. Calculations are performed in the scratch space allocated
by PDC_TMP
which is also on the lustre file system and is not node local storage.
The path to this disk space is provided by the $PDC_TMP
variable, used by
the process.scratch
directive in the dardel
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 dardel
A NAISS compute allocation should also be supplied using the --project
parameter.
Workflow organization
The workflows in this folder manage the execution of your analyses from beginning to end.
workflow/
| - .github/ Github data such as actions to run
| - assets/ Workflow assets such as test samplesheets
| - bin/ Custom workflow scripts
| - configs/ Configuration files that govern workflow execution
| - dockerfiles/ 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
| - tests/ Workflow tests
| - main.nf The primary analysis script
| - nextflow.config General Nextflow configuration
\ - modules.json nf-core file which tracks modules/subworkflows from nf-core
Version History
main @ bae13f1 (earliest) Created 23rd Aug 2024 at 14:16 by Mahesh Binzer-Panchal
Add function explanations (#114)
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Created: 23rd Aug 2024 at 14:16
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