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Commit 94d8234b authored by BIOPZ-Katsantoni Maria's avatar BIOPZ-Katsantoni Maria
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Fixed incompatibility of md5s with the multiqc merge

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1 merge request!50Polya trimming correction
Pipeline #10511 passed
...@@ -39,7 +39,7 @@ This document describes the individual rules of the pipeline for information pur ...@@ -39,7 +39,7 @@ This document describes the individual rules of the pipeline for information pur
## Detailed description of steps ## Detailed description of steps
The pipeline consists of three snakefiles: A main Snakefile and an individual Snakefile for each sequencing mode (single-end and paired-end), as parameters to individual tools differ between the sequencing modes. The main Snakefile contains some general rules for the creation of indices, rules that are applicable to both sequencing modes, and rules that deal with summary steps and combining results across samples of the run. The pipeline consists of three snakefiles: A main Snakefile and an individual Snakefile for each sequencing mode (single-end and paired-end), as parameters to individual tools differ between the sequencing modes. The main Snakefile contains some general rules for the creation of indices, rules that are applicable to both sequencing modes, and rules that deal with summary steps and combining results across samples of the run.
Individual rules of the pipeline are described briefly, and links to the respective software manuals are given. If parameters can be influenced by the user (via the samples table) they are also described. Individual rules of the pipeline are described briefly, and links to the respective software manuals are given. If parameters can be influenced by the user (via the samples table) they are also described.
Description of paired- and single-end rules are combined, only differences are highlighted. Description of paired and single-end rules are combined, only differences are highlighted.
### General ### General
...@@ -74,9 +74,10 @@ soft_clip | "Local": standard local alignment with soft-clipping allowed. "EndTo ...@@ -74,9 +74,10 @@ soft_clip | "Local": standard local alignment with soft-clipping allowed. "EndTo
pass_mode | "None": 1-pass mapping; "Basic": basic 2-pass mapping, with all 1st pass junctions inserted into the genome indices on the fly; for star mapping (type=STRING) pass_mode | "None": 1-pass mapping; "Basic": basic 2-pass mapping, with all 1st pass junctions inserted into the genome indices on the fly; for star mapping (type=STRING)
libtype | "A": automatically infer. For more info see [salmon manual](https://salmon.readthedocs.io/en/latest/salmon.html) (type=STRING) libtype | "A": automatically infer. For more info see [salmon manual](https://salmon.readthedocs.io/en/latest/salmon.html) (type=STRING)
kallisto_directionality | "--fr-stranded":Strand specific reads, first read forward. "--rf-stranded": Strand specific reads, first read reverse; for kallisto (type=STRING) kallisto_directionality | "--fr-stranded":Strand specific reads, first read forward. "--rf-stranded": Strand specific reads, first read reverse; for kallisto (type=STRING)
fq1_polya | stretch of As or Ts, depending on read orientation; for cutadapt (type=STRING) fq1_polya3p | stretch of As or Ts, depending on read orientation, trimmed from the 3' end of the read; for cutadapt (type=STRING)
fq2_polya | stretch of As or Ts, depending on read orientation; for cutadapt (type=STRING) fq1_polya5p | stretch of As or Ts, depending on read orientation, trimmed from the 5' end of the read; for cutadapt (type=STRING)
fq2_polya3p| stretch of As or Ts, depending on read orientation, trimmed from the 3' end of the read; for cutadapt (type=STRING)
fq2_polya5p| stretch of As or Ts, depending on read orientation, trimmed from the 5' end of the read; for cutadapt (type=STRING)
#### create log directories #### create log directories
Currently not implemented as Snakemake rule, but general statement. Currently not implemented as Snakemake rule, but general statement.
...@@ -260,33 +261,26 @@ Creates an interactive report after the pipeline is finished. [MultiQC](https:// ...@@ -260,33 +261,26 @@ Creates an interactive report after the pipeline is finished. [MultiQC](https://
**Output:** fastq files with adapters removed, reads shorter than 10nt will be discarded. **Output:** fastq files with adapters removed, reads shorter than 10nt will be discarded.
**Arguments not influencable by user:** **Non-customisable arguments:**
-e 0.1 maximum error-rate of 10% -e 0.1 maximum error-rate of 10%
-j 8 use 8 threads -j 8 use 8 threads
-m 10 Discard processed reads that are shorter than 10 -m 10 Discard processed reads that are shorter than 10
-n 3 search for all the given adapter sequences repeatedly, either until no adapter match was found or until 3 rounds have been performed. -n 2 search for all the given adapter sequences repeatedly, either until no adapter match was found or until 2 rounds have been performed.
*paired end:* *paired end:*
--pair-filter=both filtering criteria must apply to both reads in order for a read pair to be discarded --pair-filter=any filtering criteria must apply to any of the two reads in order for a read pair to be discarded
*single end:*
-O 1 minimal overlap of 1
#### (pe_)remove_polya_cutadapt #### (pe_)remove_polya_cutadapt
Here, [Cutadapt](https://cutadapt.readthedocs.io/en/stable/)t is used to remove poly(A) tails. Here, [Cutadapt](https://cutadapt.readthedocs.io/en/stable/) is used to remove poly(A) tails.
**Input:** fastq reads **Input:** fastq reads
**Parameters:** Adapters to be removed, specified by user in the columns 'fq1_polya', 'fq2_polya', respectively. **Parameters:** Adapters to be removed, specified by user in the columns 'fq1_polya', 'fq2_polya', respectively.
**Output:** fastq files with poly(A) tails removed, reads shorter than 10nt will be discarded. **Output:** fastq files with poly(A) tails removed, reads shorter than 10nt will be discarded.
**Arguments like in remove_adapters_cutadapt and additionally:** **Arguments similar to remove_adapters_cutadapt and additionally:**
--match-read-wildcards This option is used to allow matching wildcard characters also within reads, because if no tail should be trimmed "XXXXXX" is specified in the samples table, which doesn't match any nucleotides, and thus nothing will be done here. -n 1 search for all the given adapter sequences repeatedly, either until no adapter match was found or until 1 round has been performed.
-n 2 search for all the given adapter sequences repeatedly, either until no adapter match was found or until 2 rounds have been performed.
-q 6 trim low-quality 3'ends with a cutoff of 6 nucleotides
*paired end:* *paired end:*
--pair-filter=both filtering criteria must apply to both reads in order for a read pair to be discarded --pair-filter=any filtering criteria must apply to both reads in order for a read pair to be discarded
*single end:* *single end:*
-O 1 minimal overlap of 1 -O 1 minimal overlap of 1
...@@ -318,8 +312,6 @@ Spliced Transcripts Alignment to a Reference; Read the [Publication](https://www ...@@ -318,8 +312,6 @@ Spliced Transcripts Alignment to a Reference; Read the [Publication](https://www
*Same for single- and paired-end.* *Same for single- and paired-end.*
#### (pe_)quantification_salmon #### (pe_)quantification_salmon
[Salmon](https://salmon.readthedocs.io/en/latest/salmon.html) is a tool for wicked-fast transcript quantification from RNA-seq data. [Salmon](https://salmon.readthedocs.io/en/latest/salmon.html) is a tool for wicked-fast transcript quantification from RNA-seq data.
...@@ -358,4 +350,3 @@ Spliced Transcripts Alignment to a Reference; Read the [Publication](https://www ...@@ -358,4 +350,3 @@ Spliced Transcripts Alignment to a Reference; Read the [Publication](https://www
* -l: fragment length, user specified as `mean` * -l: fragment length, user specified as `mean`
* -s: fragment length SD, user specified as `sd` * -s: fragment length SD, user specified as `sd`
...@@ -19,7 +19,7 @@ ea36f062eedc7f54ceffea2b635a25a8 results/star_indexes/homo_sapiens/75/STAR_inde ...@@ -19,7 +19,7 @@ ea36f062eedc7f54ceffea2b635a25a8 results/star_indexes/homo_sapiens/75/STAR_inde
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500dd49da40b16799aba62aa5cf239ba results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/synthetic_10_reads_paired_synthetic_10_reads_paired.remove_polya_mate1.fastq 500dd49da40b16799aba62aa5cf239ba results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/synthetic_10_reads_paired_synthetic_10_reads_paired.remove_polya_mate1.fastq
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f551ff091e920357ec0a76807cb51dba results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/mate1_fastqc/synthetic.mate_1_fastqc/fastqc_data.txt f551ff091e920357ec0a76807cb51dba results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/mate1_fastqc/synthetic.mate_1_fastqc/fastqc_data.txt
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...@@ -45,8 +45,8 @@ b28aac49f537b8cba364b6422458ad28 results/samples/synthetic_10_reads_paired_synt ...@@ -45,8 +45,8 @@ b28aac49f537b8cba364b6422458ad28 results/samples/synthetic_10_reads_paired_synt
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5b950b5dfe3c7407e9aac153db330a38 results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/mate2_fastqc/synthetic.mate_2_fastqc/Images/sequence_length_distribution.png 5b950b5dfe3c7407e9aac153db330a38 results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/mate2_fastqc/synthetic.mate_2_fastqc/Images/sequence_length_distribution.png
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c77480e0235761f2d7f80dbceb2e2806 results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/salmon_quant/synthetic_10_reads_paired_synthetic_10_reads_paired/lib_format_counts.json c77480e0235761f2d7f80dbceb2e2806 results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/salmon_quant/synthetic_10_reads_paired_synthetic_10_reads_paired/lib_format_counts.json
989d6ee63b728fced9ec0249735ab83d results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/salmon_quant/synthetic_10_reads_paired_synthetic_10_reads_paired/aux_info/ambig_info.tsv 989d6ee63b728fced9ec0249735ab83d results/samples/synthetic_10_reads_paired_synthetic_10_reads_paired/salmon_quant/synthetic_10_reads_paired_synthetic_10_reads_paired/aux_info/ambig_info.tsv
...@@ -78,10 +78,10 @@ e72f5d798c99272f8c0166dc77247db1 results/samples/synthetic_10_reads_mate_1_synt ...@@ -78,10 +78,10 @@ e72f5d798c99272f8c0166dc77247db1 results/samples/synthetic_10_reads_mate_1_synt
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...@@ -89,9 +89,9 @@ bcccf679a8c083d01527514c9f5680a0 results/samples/synthetic_10_reads_mate_1_synt ...@@ -89,9 +89,9 @@ bcccf679a8c083d01527514c9f5680a0 results/samples/synthetic_10_reads_mate_1_synt
3ce47cb1d62482c5d62337751d7e8552 results/transcriptome/homo_sapiens/transcriptome.fa 3ce47cb1d62482c5d62337751d7e8552 results/transcriptome/homo_sapiens/transcriptome.fa
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\ No newline at end of file
...@@ -7,7 +7,7 @@ cleanup () { ...@@ -7,7 +7,7 @@ cleanup () {
rm -rf .java/ rm -rf .java/
rm -rf .snakemake/ rm -rf .snakemake/
rm -rf logs/ rm -rf logs/
rm -rf results/ # rm -rf results/
cd $user_dir cd $user_dir
echo "Exit status: $rc" echo "Exit status: $rc"
} }
......
...@@ -6,6 +6,7 @@ cleanup () { ...@@ -6,6 +6,7 @@ cleanup () {
rm -rf .snakemake/ rm -rf .snakemake/
rm -rf config.yaml rm -rf config.yaml
rm -rf samples.tsv rm -rf samples.tsv
rm -rf logs
cd $user_dir cd $user_dir
echo "Exit status: $rc" echo "Exit status: $rc"
} }
......
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