Nextflow pipeline that assembles mitochondria scaffolds using MITObim, checks the scaffold by mapping reads to it using bwa mem, and calling variants in freebayes to obtain a consensus sequence. Workflow is based on a Mozes Blom
pipeline.
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Install
nextflow
(version >= 19.04)
InstallConda
(version >= 4.10)
Installmitobim
Installmira
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Download git clone of this repository:
git clone https://github.com/FilipThorn/nf_mito-mania
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Edit nextflow.config file:
mitobim = "/PATH/TO/MITObim.pl" #path to MITObim script mitobimRef = "/PATH/TO/lycPyr_mtDNA.fa" #refernce for mitobim and mira ref_strain = "lycPyr_mtDNA" #name of refernce for mitobim and mira mira = "/PATH/TO/mira_4.0.2_linux-gnu_x86_64_static/bin/mira" #path to mira mira_dir = "/PATH/TO/mira_4.0.2_linux-gnu_x86_64_static/bin/" #path to mira dir
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Run Mitomania workflow:
nextflow run mitomania.nf --reads_MB /PATH/TO/Indivxxx_L001_U.fastq.gz --reads_PE '/PATH/TO/*_R{1,2}.fastq.gz' --reads_SE '/PATH/TO/*_U.fastq.gz' --outdir /PATH/TO/RESULTS
Currently requires both paired and unpaired reads to run as well as four seperate libraries per individual. Updates to generalize the script are ongoing.
Input file structure
usr:~data/$ ls
Indivxxx_L001_U.fastq.gz Indivxxx_L001_R2.fastq.gz Indivxxx_L001_R1.fastq.gz
Indivxxx_L002_U.fastq.gz Indivxxx_L002_R2.fastq.gz Indivxxx_L002_R1.fastq.gz
Indivxxx_L003_U.fastq.gz Indivxxx_L003_R2.fastq.gz Indivxxx_L003_R1.fastq.gz
Indivxxx_L004_U.fastq.gz Indivxxx_L004_R2.fastq.gz Indivxxx_L004_R1.fastq.gz
Use of a HPC is recomended. Create a nextflow config profile that matches your cluster set-up profile