Remove sample specific options and use them as rule specific.

pipeline_documentation.md

@@ -116,9 +116,6 @@ gtf_filtered | Required for [Salmon](#third-party-software-used). Path to filter
 genome | Required for [STAR](#third-party-software-used). Path to genome `.fa` file. File needs to be in subdirectory corresponding to `organism` field. Example: `/path/to/GRCh38/genome.fa` | `str`
 sd | Required for [kallisto](#third-party-software-used) and [Salmon](#third-party-software-used), but only for single-end libraries. Estimated standard deviation of fragment length distribution. Can be assessed from, e.g., BioAnalyzer profiles. Value ignored for paired-end libraries. | `int`
 mean | Required for [kallisto](#third-party-software-used) and [Salmon](#third-party-software-used), but only for single-end libraries. Estimated mean of fragment length distribution. Can be assessed, e.g., from BioAnalyzer profiles. Value ignored for paired-end libraries. | `int`
-multimappers | Required for [STAR](#third-party-software-used). Maximum number of multiple alignments allowed for a read; if exceeded, the read is considered unmapped. | `int`
-soft_clip | Required for [STAR](#third-party-software-used). One of `Local` (standard local alignment with soft-clipping allowed) or `EndToEnd` (force end-to-end read alignment, do not soft-clip). | `str`
-pass_mode | Required for [STAR](#third-party-software-used). One of `None` (1-pass mapping) or `Basic` (basic 2-pass mapping, with all 1st-pass junctions inserted into the genome indices on the fly). | `str`
 libtype | Required for [Salmon](#third-party-software-used), and, after internal conversion, for [kallisto](#third-party-software-used) and [ALFA](#third-party-software-used) . See [Salmon manual][docs-salmon] for allowed values.    **WARNING**: do *NOT* use `A` to automatically infer the salmon library type, this will cause kallisto and ALFA to fail.  | `str`
 fq1_polya3p | Required for [Cutadapt](#third-party-software-used). Stretch of `A`s or `T`s, depending on read orientation. Trimmed from the 3' end of the read. Use value such as `XXXXXXXXXXXXXXX` if no poly(A) stretch present or if no trimming is desired. | `str`
 fq1_polya5p | Required for [Cutadapt](#third-party-software-used). Stretch of `A`s or `T`s, depending on read orientation. Trimmed from the 5' end of the read. Use value such as `XXXXXXXXXXXXXXX` if no poly(A) stretch present or if no trimming is desired. | `str`
@@ -612,13 +609,12 @@ Align short reads to reference genome and/or transcriptome with
     [**remove_polya_cutadapt**](#remove_polya_cutadapt)
   - Index; from [**create_index_star**](#create_index_star)
 - **Parameters**
-  - **samples.tsv**
-    - `--outFilterMultimapNmax`: maximum number of multiple alignments allowed; if exceeded, read is considered unmapped; specify in sample table column `multimappers`
-    - `--alignEndsType`: one of `Local` (standard local alignment with soft-clipping allowed) or `EndToEnd` (force end-to-end read alignment, do not soft-clip); specify in sample table column `soft_clip`
-    - `--twopassMode`: one of `None` (1-pass mapping) or `Basic` (basic 2-pass mapping, with all 1st-pass junctions inserted into the genome indices on the fly); specify in sample table column `pass_mode`
   - **rule_config.yaml**
     - `--outFilterMultimapScoreRange=0`: the score range below the maximum score for multimapping alignments (default 1)
     - `--outFilterType=BySJout`: reduces the number of ”spurious” junctions
+    - `--alignEndsType`: one of `Local` (standard local alignment with soft-clipping allowed) or `EndToEnd` (force end-to-end read alignment, do not soft-clip); specify in sample table column `soft_clip`
+    - `--twopassMode`: one of `None` (1-pass mapping) or `Basic` (basic 2-pass mapping, with all 1st-pass junctions inserted into the genome indices on the fly); specify in sample table column `pass_mode`
+    - `--outFilterMultimapNmax`: maximum number of multiple alignments allowed; if exceeded, read is considered unmapped; specify in sample table column `multimappers`
 - **Output**
   - Aligned reads file (`.bam`); used in
     [**calculate_TIN_scores**](#calculate_TIN_scores),

tests/input_files/rule_config.yaml

@@ -160,6 +160,12 @@ map_genome_star:
     # "SJ.out.tab" (default 'Normal', ZARP recommends 'BySJout', as this
     # reduces the number of spurious junctions )
     --outFilterType: 'BySJout'
+    # type of read ends alignment: force end-to-end read alignment, do not soft-clip
+    --alignEndsType: 'EndToEnd'
+    #  extract junctions, insert them into the genome index and re-map reads in a 2nd mapping pass
+    --twopassMode: Basic
+    # alignments (all of them) will be output only if the read maps to no more loci than 10
+    --outFilterMultimapNmax: '10'
 
 pe_map_genome_star:
     # The score range below the maximum score for multimapping alignments
@@ -169,6 +175,12 @@ pe_map_genome_star:
     # "SJ.out.tab" (default 'Normal', ZARP recommends 'BySJout', as this
     # reduces the number of spurious junctions )
     --outFilterType: 'BySJout'
+    # type of read ends alignment: force end-to-end read alignment, do not soft-clip
+    --alignEndsType: 'EndToEnd'
+    #  extract junctions, insert them into the genome index and re-map reads in a 2nd mapping pass
+    --twopassMode: Basic
+    # alignments (all of them) will be output only if the read maps to no more loci than 10
+    --outFilterMultimapNmax: '10'
 
 quantification_salmon:
     # Correct for sequence specific biases; cf.

tests/input_files/samples.multiple_lanes.tsv

-sample	seqmode	fq1	fq2	index_size	kmer	fq1_3p	fq1_5p	fq2_3p	fq2_5p	organism	gtf	genome	sd	mean	multimappers	soft_clip	pass_mode	libtype	fq1_polya_3p	fq1_polya_5p	fq2_polya_3p	fq2_polya_5p
-synthetic_10_reads_paired_synthetic_10_reads_paired	pe	../input_files/pe_lane1/synthetic_split_lane1.mate_1.fastq.gz	../input_files/pe_lane1/synthetic_split_lane1.mate_2.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	10	EndToEnd	None	ISF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	TTTTTTTTTTTTTTT
-synthetic_10_reads_paired_synthetic_10_reads_paired	pe	../input_files/pe_lane2/synthetic_split_lane2.mate_1.fastq.gz	../input_files/pe_lane2/synthetic_split_lane2.mate_2.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	10	EndToEnd	None	ISF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	TTTTTTTTTTTTTTT
-synthetic_10_reads_mate_1_synthetic_10_reads_mate_1	se	../input_files/se_lane1/synthetic_split_lane1.mate_1.fastq.gz	XXXXXXXXXXXXXXX	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	10	EndToEnd	None	SF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX
-synthetic_10_reads_mate_1_synthetic_10_reads_mate_1	se	../input_files/se_lane2/synthetic_split_lane2.mate_1.fastq.gz	XXXXXXXXXXXXXXX	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	10	EndToEnd	None	SF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX
+sample	seqmode	fq1	fq2	index_size	kmer	fq1_3p	fq1_5p	fq2_3p	fq2_5p	organism	gtf	genome	sd	mean	libtype	fq1_polya_3p	fq1_polya_5p	fq2_polya_3p	fq2_polya_5p
+synthetic_10_reads_paired_synthetic_10_reads_paired	pe	../input_files/pe_lane1/synthetic_split_lane1.mate_1.fastq.gz	../input_files/pe_lane1/synthetic_split_lane1.mate_2.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	ISF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	TTTTTTTTTTTTTTT
+synthetic_10_reads_paired_synthetic_10_reads_paired	pe	../input_files/pe_lane2/synthetic_split_lane2.mate_1.fastq.gz	../input_files/pe_lane2/synthetic_split_lane2.mate_2.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	ISF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	TTTTTTTTTTTTTTT
+synthetic_10_reads_mate_1_synthetic_10_reads_mate_1	se	../input_files/se_lane1/synthetic_split_lane1.mate_1.fastq.gz	XXXXXXXXXXXXXXX	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	SF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX
+synthetic_10_reads_mate_1_synthetic_10_reads_mate_1	se	../input_files/se_lane2/synthetic_split_lane2.mate_1.fastq.gz	XXXXXXXXXXXXXXX	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	SF	AAAAAAAAAAAAAAA	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX	XXXXXXXXXXXXXXX

tests/input_files/samples.tsv

-sample	seqmode	fq1	index_size	kmer	fq1_3p	fq1_5p	organism	gtf	genome	sd	mean	multimappers	soft_clip	pass_mode	libtype	fq1_polya_3p	fq1_polya_5p	fq2	fq2_3p	fq2_5p	fq2_polya_3p	fq2_polya_5p
-synthetic_10_reads_paired_synthetic_10_reads_paired	pe	../input_files/project1/synthetic.mate_1.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	10	EndToEnd	None	ISF	AAAAAAAAAAAAAAAAA	XXXXXXXXXXXXXXXXX	../input_files/project1/synthetic.mate_2.fastq.gz	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	XXXXXXXXXXXXXXXXX	TTTTTTTTTTTTTTTTT
-synthetic_10_reads_mate_1_synthetic_10_reads_mate_1	se	../input_files/project2/synthetic.mate_1.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	10	EndToEnd	None	SF	AAAAAAAAAAAAAAAAA	XXXXXXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX
+sample	seqmode	fq1	index_size	kmer	fq1_3p	fq1_5p	organism	gtf	genome	sd	mean	libtype	fq1_polya_3p	fq1_polya_5p	fq2	fq2_3p	fq2_5p	fq2_polya_3p	fq2_polya_5p
+synthetic_10_reads_paired_synthetic_10_reads_paired	pe	../input_files/project1/synthetic.mate_1.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	ISF	AAAAAAAAAAAAAAAAA	XXXXXXXXXXXXXXXXX	../input_files/project1/synthetic.mate_2.fastq.gz	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	XXXXXXXXXXXXXXXXX	TTTTTTTTTTTTTTTTT
+synthetic_10_reads_mate_1_synthetic_10_reads_mate_1	se	../input_files/project2/synthetic.mate_1.fastq.gz	75	31	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/annotation.gtf	../input_files/homo_sapiens/genome.fa	100	250	SF	AAAAAAAAAAAAAAAAA	XXXXXXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX	XXXXXXXXXXXXX
\ No newline at end of file

tests/input_files/samples_alfa.tsv

-sample	seqmode	fq1	index_size	kmer	fq2	fq1_3p	fq1_5p	fq2_3p	fq2_5p	organism	gtf	genome	sd	mean	multimappers	soft_clip	pass_mode	libtype	fq1_polya	fq2_polya
-paired_end_R1_on_plus_sense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	GATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	10	EndToEnd	None	ISF	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT
-paired_end_R1_on_plus_antisense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	10	EndToEnd	None	ISR	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT
-paired_end_R1_on_minus_sense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	10	EndToEnd	None	ISF	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT
-paired_end_R1_on_minus_antisense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	10	EndToEnd	None	ISR	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT
+sample	seqmode	fq1	index_size	kmer	fq2	fq1_3p	fq1_5p	fq2_3p	fq2_5p	organism	gtf	genome	sd	mean	libtype	fq1_polya	fq2_polya
+paired_end_R1_on_plus_sense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	GATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	ISF	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT
+paired_end_R1_on_plus_antisense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	ISR	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT
+paired_end_R1_on_minus_sense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	ISF	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT
+paired_end_R1_on_minus_antisense	pe	XXXXXXXXXXXXX	75	31	XXXXXXXXXXXXX	AGATCGGAAGAGCACA	XXXXXXXXXXXXX	AGATCGGAAGAGCGT	XXXXXXXXXXXXX	homo_sapiens	../input_files/homo_sapiens/quick_start.gtf	XXXXXXXXXXXXX	100	250	ISR	AAAAAAAAAAAAAAAAA	TTTTTTTTTTTTTTTTT

workflow/rules/paired_end.snakefile.smk

@@ -256,36 +256,18 @@ rule pe_map_genome_star:
             "{sample}",
             "map_genome",
             "{sample}.pe."),
-        multimappers = lambda wildcards:
-            get_sample(
-                'multimappers',
-                search_id='index',
-                search_value=wildcards.sample),
-        soft_clip = lambda wildcards:
-            get_sample(
-                'soft_clip',
-                search_id='index',
-                search_value=wildcards.sample),
-        pass_mode = lambda wildcards:
-            get_sample(
-                'pass_mode',
-                search_id='index',
-                search_value=wildcards.sample),
         additional_params = parse_rule_config(
             rule_config,
             current_rule=current_rule,
             immutable=(
-                '--twopassMode',
                 '--genomeDir',
                 '--readFilesIn',
                 '--readFilesCommand',
-                '--outFilterMultimapNmax',
                 '--outFileNamePrefix',
                 '--outSAMattributes',
                 '--outStd',
                 '--outSAMtype',
                 '--outSAMattrRGline',
-                '--alignEndsType',
                 )
             )
 
@@ -306,18 +288,15 @@ rule pe_map_genome_star:
 
     shell:
         "(STAR \
-        --twopassMode {params.pass_mode} \
         --runThreadN {threads} \
         --genomeDir {params.index} \
         --readFilesIn {input.reads1} {input.reads2} \
         --readFilesCommand zcat \
-        --outFilterMultimapNmax {params.multimappers} \
         --outFileNamePrefix {params.outFileNamePrefix} \
         --outSAMattributes All \
         --outStd BAM_SortedByCoordinate \
         --outSAMtype BAM SortedByCoordinate \
         --outSAMattrRGline ID:rnaseq_pipeline SM:{params.sample_id} \
-        --alignEndsType {params.soft_clip} \
         {params.additional_params} \
         > {output.bam};) \
         2> {log.stderr}"

workflow/rules/single_end.snakefile.smk

@@ -200,36 +200,18 @@ rule map_genome_star:
             "{sample}",
             "map_genome",
             "{sample}.se."),
-        multimappers = lambda wildcards:
-            get_sample(
-                'multimappers',
-                search_id='index',
-                search_value=wildcards.sample),
-        soft_clip = lambda wildcards:
-            get_sample(
-                'soft_clip',
-                search_id='index',
-                search_value=wildcards.sample),
-        pass_mode = lambda wildcards:
-            get_sample(
-                'pass_mode',
-                search_id='index',
-                search_value=wildcards.sample),
         additional_params = parse_rule_config(
             rule_config,
             current_rule=current_rule,
             immutable=(
-                '--twopassMode',
                 '--genomeDir',
                 '--readFilesIn',
                 '--readFilesCommand',
-                '--outFilterMultimapNmax',
                 '--outFileNamePrefix',
                 '--outSAMattributes',
                 '--outStd',
                 '--outSAMtype',
                 '--outSAMattrRGline',
-                '--alignEndsType',
                 )
             )
 
@@ -250,18 +232,15 @@ rule map_genome_star:
 
     shell:
         "(STAR \
-        --twopassMode {params.pass_mode} \
         --runThreadN {threads} \
         --genomeDir {params.index} \
         --readFilesIn {input.reads} \
         --readFilesCommand zcat \
-        --outFilterMultimapNmax {params.multimappers} \
         --outFileNamePrefix {params.outFileNamePrefix} \
         --outSAMattributes All \
         --outStd BAM_SortedByCoordinate \
         --outSAMtype BAM SortedByCoordinate \
         --outSAMattrRGline ID:rnaseq_pipeline SM:{params.sample_id} \
-        --alignEndsType {params.soft_clip} \
         {params.additional_params} \
         > {output.bam};) \
         2> {log.stderr}"