diff --git a/bin/calculate_rrna_mtdna.sh b/bin/calculate_rrna_mtdna.sh old mode 100644 new mode 100755 index c677ddd..be99562 --- a/bin/calculate_rrna_mtdna.sh +++ b/bin/calculate_rrna_mtdna.sh @@ -45,6 +45,24 @@ echo -e "Number of ribosomal RNA reads in uniquely mapped reads,$rrna" >> $outfi perc_rrna=$(awk -v r="$rrna" -v u="$uniquely_mapped" 'BEGIN {printf "%.4f", (r/u)}') echo -e "Percentage of rRNA reads (of uniquely mapped reads),$perc_rrna" >> $outfile +# Count rRNA in multimapped primary alignments +featureCounts -t "${grep_rrna}" -a ${ref_gtf} -o feat_counts_rRNA_mmpa.txt multimapped_primealign.bam +rrna_mmpa=$(cat feat_counts_rRNA_mmpa.txt.summary | grep -E "Assigned" | awk '{print $2}') +echo -e "rRNA counts in Multimapped reads (primary alignment),$rrna_mmpa" >> $outfile + +# Percentage of rRNA in multimapped primary alignments +perc_rrna_mmpa=$(awk -v r="$rrna_mmpa" -v tot="$total_mmpa" 'BEGIN {printf "%.4f", (r/tot)}') +echo -e "Percentage of rRNA in multimapped reads (primary alignment),$perc_rrna_mmpa" >> $outfile + +# Count rRNA in multimapped all alignments +featureCounts -t "${grep_rrna}" -a ${ref_gtf} -o feat_counts_rRNA_mmaa.txt multimapped_allalign.bam +rrna_mmaa=$(cat feat_counts_rRNA_mmaa.txt.summary | grep -E "Assigned" | awk '{print $2}') +echo -e "rRNA counts in Multimapped reads (all alignments),$rrna_mmaa" >> $outfile + +# Percentage of rRNA in multimapped all alignments +perc_rrna_mmaa=$(awk -v r="$rrna_mmaa" -v tot="$total_mmaa" 'BEGIN {printf "%.4f", (r/tot)}') +echo -e "Percentage of rRNA in multimapped reads (all alignments),$perc_rrna_mmaa" >> $outfile + # ------------------------------------------------------------------ # Calculate mtDNA metrics diff --git a/modules/local/custom/demultiplex/parsebio_demultiplex/main.nf b/modules/local/custom/demultiplex/parsebio_demultiplex/main.nf index fc93d47..1156e57 100644 --- a/modules/local/custom/demultiplex/parsebio_demultiplex/main.nf +++ b/modules/local/custom/demultiplex/parsebio_demultiplex/main.nf @@ -10,12 +10,11 @@ process PARSEBIO_CUSTOM_DEMUX { tuple val(meta), path(fastq_cDNA), path(fastq_BC_UMI), path(fastq_indices), path(input_file) output: - tuple val(meta), path("*group*_R1*"), path("*group*_R2*"), path(input_file), emit: splitted_files + tuple val(meta), path("*group*_R1*"), path("*group*_R2*"), path(fastq_indices), path(input_file), emit: splitted_files script: """ echo "\n\n================== Parse Biosciences: Custom Demultiplexing ==================" - echo "Conda environment: \$CONDA_DEFAULT_ENV" echo "Processing sample: ${meta}" echo "Fastq files: ${fastq_cDNA}, ${fastq_BC_UMI}" echo "Group id: ${meta.id}, wells (rt): ${meta.rt}" diff --git a/modules/local/tools/cellbender/main.nf b/modules/local/tools/cellbender/main.nf index f02c7a5..ba876f6 100644 --- a/modules/local/tools/cellbender/main.nf +++ b/modules/local/tools/cellbender/main.nf @@ -3,10 +3,7 @@ process CELLBENDER { tag "${meta.id}" label 'process_high_memory' - container "${ task.ext.use_gpu ? 'us.gcr.io/broad-dsde-methods/cellbender:0.3.2' : - workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://community-cr-prod.seqera.io/docker/registry/v2/blobs/sha256/eb/ebcf140f995f79fcad5c17783622e000550ff6f171771f9fc4233484ee6f63cf/data': - 'community.wave.seqera.io/library/cellbender_webcolors:156d413fdfc16cdb' }" + container "oras://community.wave.seqera.io/library/cellbender:0.3.2--4f86af6695399b4f" input: tuple val(meta), path(mapping_files) diff --git a/modules/local/tools/cutadapt/main.nf b/modules/local/tools/cutadapt/main.nf index da0dd62..aafdf0c 100644 --- a/modules/local/tools/cutadapt/main.nf +++ b/modules/local/tools/cutadapt/main.nf @@ -4,9 +4,7 @@ process RM_VARBASES { conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://community-cr-prod.seqera.io/docker/registry/v2/blobs/sha256/17/1758869538eb8e658077cc14cd7a4e76fd9b6d73d3a68f85a70bf292e39e27c5/data' : - 'community.wave.seqera.io/library/cutadapt:5.0--991bbd2e184b7014' }" + container "oras://community.wave.seqera.io/library/cutadapt:5.2--5505472a18e9cce0" input: tuple val(meta), path(fastq_cDNA), path(fastq_BC_UMI), path(fastq_indices), path(input_file) diff --git a/modules/local/tools/fastp/main.nf b/modules/local/tools/fastp/main.nf index 0bfb51a..16fbce4 100644 --- a/modules/local/tools/fastp/main.nf +++ b/modules/local/tools/fastp/main.nf @@ -5,9 +5,7 @@ process FASTP { debug true conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://community-cr-prod.seqera.io/docker/registry/v2/blobs/sha256/88/889a182b8066804f4799f3808a5813ad601381a8a0e3baa4ab8d73e739b97001/data' : - 'community.wave.seqera.io/library/fastp:0.24.0--62c97b06e8447690' }" + container "oras://community.wave.seqera.io/library/fastp:0.24.0--0397de619771c7ae" input: tuple val(meta), path(fastq_cDNA), path(fastq_BC_UMI), path(fastq_indices), path(input_file) diff --git a/modules/local/tools/fastqc/main.nf b/modules/local/tools/fastqc/main.nf index 5ac5990..d976b98 100644 --- a/modules/local/tools/fastqc/main.nf +++ b/modules/local/tools/fastqc/main.nf @@ -4,9 +4,7 @@ process FASTQC { label 'process_low' conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/fastqc:0.12.1--hdfd78af_0' : - 'quay.io/biocontainers/fastqc:0.12.1--hdfd78af_0' }" + container "oras://community.wave.seqera.io/library/fastqc:0.12.1--104d26ddd9519960" input: tuple val(meta), path(fastq_cDNA), path(fastq_BC_UMI), path(fastq_indices), path(input_file) diff --git a/modules/local/tools/kraken/kraken_classify/main.nf b/modules/local/tools/kraken/kraken_classify/main.nf index 85c516b..2819b37 100644 --- a/modules/local/tools/kraken/kraken_classify/main.nf +++ b/modules/local/tools/kraken/kraken_classify/main.nf @@ -8,9 +8,7 @@ process KRAKEN { tuple val(meta), path(mapping_files) conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://community-cr-prod.seqera.io/docker/registry/v2/blobs/sha256/29/29ed8f68315625eca61a3de9fcb7b8739fe8da23f5779eda3792b9d276aa3b8f/data' : - 'community.wave.seqera.io/library/kraken2_coreutils_pigz:45764814c4bb5bf3' }" + container "oras://community.wave.seqera.io/library/kraken2_samtools_coreutils_pigz:8961943c277652a6" output: path("*") diff --git a/modules/local/tools/krona/main.nf b/modules/local/tools/krona/main.nf index 47aeec5..f3ecfc4 100644 --- a/modules/local/tools/krona/main.nf +++ b/modules/local/tools/krona/main.nf @@ -4,9 +4,7 @@ process KRONA { conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/krona:2.8--pl5262hdfd78af_2' : - 'quay.io/biocontainers/krona:2.8--pl5262hdfd78af_2' }" + container "oras://community.wave.seqera.io/library/krona:2.8--8fc5aef4acd456a6" input: val(trigger) diff --git a/modules/local/tools/multiqc/main.nf b/modules/local/tools/multiqc/main.nf index aa7baa7..b0dca06 100644 --- a/modules/local/tools/multiqc/main.nf +++ b/modules/local/tools/multiqc/main.nf @@ -4,9 +4,7 @@ process MULTIQC { conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/multiqc:1.30--pyhdfd78af_1' : - 'quay.io/biocontainers/multiqc:1.30--pyhdfd78af_1' }" + container "oras://community.wave.seqera.io/library/multiqc:1.30--d3e586af7b974fba" input: val(trigger) diff --git a/modules/local/tools/salmon_alevin/alevin-fry/main.nf b/modules/local/tools/salmon_alevin/alevin-fry/main.nf index 3a8c381..f2f34bc 100644 --- a/modules/local/tools/salmon_alevin/alevin-fry/main.nf +++ b/modules/local/tools/salmon_alevin/alevin-fry/main.nf @@ -5,9 +5,7 @@ process ALEVIN_FRY { conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/simpleaf:0.19.4--ha6fb395_0': - 'quay.io/biocontainers/simpleaf:0.19.4--ha6fb395_0' }" + container "oras://community.wave.seqera.io/library/alevin-fry_piscem_salmon_simpleaf_pruned:c71cfb476b003414" input: tuple val(meta), path(fastq_cDNA), path(fastq_BC_UMI), path(fastq_indices), path(input_file) diff --git a/modules/local/tools/salmon_alevin/salmon_index/main.nf b/modules/local/tools/salmon_alevin/salmon_index/main.nf index 65daf9e..f414bd3 100644 --- a/modules/local/tools/salmon_alevin/salmon_index/main.nf +++ b/modules/local/tools/salmon_alevin/salmon_index/main.nf @@ -4,9 +4,7 @@ process SALMON_INDEX { conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/simpleaf:0.19.4--ha6fb395_0': - 'quay.io/biocontainers/simpleaf:0.19.4--ha6fb395_0' }" + container "oras://community.wave.seqera.io/library/salmon:1.10.3--726401738a281398" input: tuple val(meta), path(fastq_cDNA), path(fastq_BC_UMI), path(fastq_indices), path(input_file) diff --git a/modules/local/tools/samtools/samtools_index/main.nf b/modules/local/tools/samtools/samtools_index/main.nf index 137fff5..985c0df 100644 --- a/modules/local/tools/samtools/samtools_index/main.nf +++ b/modules/local/tools/samtools/samtools_index/main.nf @@ -4,9 +4,7 @@ process SAMTOOLS_INDEX { label 'process_single' conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/samtools:1.22.1--h96c455f_0' : - 'quay.io/biocontainers/samtools:1.22.1--h96c455f_0' }" + container "oras://community.wave.seqera.io/library/samtools:1.22.1--9a10f06c24cdf05f" input: tuple val(meta), path(mapping_files) diff --git a/modules/local/tools/samtools/samtools_view/main.nf b/modules/local/tools/samtools/samtools_view/main.nf index 7ace27a..f4691f9 100644 --- a/modules/local/tools/samtools/samtools_view/main.nf +++ b/modules/local/tools/samtools/samtools_view/main.nf @@ -4,9 +4,7 @@ process SAMTOOLS_VIEW { label 'process_single' conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/samtools:1.22.1--h96c455f_0' : - 'quay.io/biocontainers/samtools:1.22.1--h96c455f_0' }" + container "oras://community.wave.seqera.io/library/samtools:1.22.1--9a10f06c24cdf05f" input: tuple val(meta), path(mapping_files) diff --git a/modules/local/tools/scirocket/scirocket_demux/main.nf b/modules/local/tools/scirocket/scirocket_demux/main.nf index e7e5e45..244fa04 100644 --- a/modules/local/tools/scirocket/scirocket_demux/main.nf +++ b/modules/local/tools/scirocket/scirocket_demux/main.nf @@ -4,8 +4,8 @@ process SCIROCKET_DEMUX { label 'process_medium' debug true - conda "${moduleDir}/environment.yml" + container "oras://community.wave.seqera.io/library/fastp_pysam_sambamba_star_pruned:ac8b1ecd993405dd" input: tuple val(meta), path(fastq_cDNA), path(fastq_BC_UMI), path(fastq_indices), path(input_file) @@ -29,7 +29,6 @@ process SCIROCKET_DEMUX { """ echo "\n\n================== DEMULTIPLEXING FASTQ FILES ==================" - echo "Conda environment: \$CONDA_DEFAULT_ENV" echo "Sample ID: ${meta.id}" echo "FASTQ cDNA: ${fastq_cDNA}" echo "FASTQ BC & UMI: ${fastq_BC_UMI}" diff --git a/modules/local/tools/star/starsolo_genome_generate/main.nf b/modules/local/tools/star/starsolo_genome_generate/main.nf index c8f5c8b..f96c7ac 100644 --- a/modules/local/tools/star/starsolo_genome_generate/main.nf +++ b/modules/local/tools/star/starsolo_genome_generate/main.nf @@ -7,9 +7,7 @@ process STARSOLO_INDEX { path ref_gtf conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://community-cr-prod.seqera.io/docker/registry/v2/blobs/sha256/26/268b4c9c6cbf8fa6606c9b7fd4fafce18bf2c931d1a809a0ce51b105ec06c89d/data' : - 'community.wave.seqera.io/library/htslib_samtools_star_gawk:ae438e9a604351a4' }" + container "oras://community.wave.seqera.io/library/htslib_samtools_star_gawk:f196f82abbbc8871" output: path("*")