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author | bgruening |
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date | Tue, 07 Jun 2016 15:47:26 -0400 |
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<tool id="ribotaper_ribosome_profiling" name="ribotaper part 3: ribosome profiling" version="0.1.0"> <requirements> <requirement type="package" version="1.3.1">ribotaper</requirement> </requirements> <stdio> <exit_code range="1:" /> </stdio> <command><![CDATA[ tar "xzvf" "$annotation_path" && Ribotaper.sh "$ribo_bam" "$rna_bam" "annotation_path" "$read_lenghts_ribo1,$read_lenghts_ribo2,$read_lenghts_ribo3" "$cutoff1,$cutoff2,$cutoff3" "\${GALAXY_SLOTS:-12}" ]]></command> <inputs> <param name="annotation_path" type="data" format="compressed_archive" label="annotation_path" help="Please run 'ribotaper part 1' to generate the archive."/> <param name="ribo_bam" type="data" format="BAM" label="ribo_bam" help="Ribo-seq alignment file in BAM format."/> <param name="rna_bam" type="data" format="BAM" label="rna_bam" help="RNA-seq alignment file in BAM format."/> <param name="read_lenghts_ribo1" type="text" value="26" label="Read length 1" help="Read length 1, which is used for P-site calculation. Default is '26' but it varies a lot in different datasets. Please run 'ribotaper part 2' to deterimine a appropriate value."/> <param name="read_lenghts_ribo2" type="text" value="28" label="Read length 2" help="Read length 2, which is used for P-site calculation. Default is '28' but it varies a lot in different datasets. Please run 'ribotaper part 2' to deterimine a appropriate value"/> <param name="read_lenghts_ribo3" type="text" value="29" label="Read length 3" help="Read length 3, which is used for P-site calculation. Default is '29' but it varies a lot in different datasets. Please run 'ribotaper part 2' to deterimine a appropriate value"/> <param name="cutoff1" type="text" value="9" label="Cutoff 1" help="Offset 1, which is used for P-sites calculation. Default is '9' but it varies a lot in different datasets. Please run 'ribotaper part 2' to deterimine a appropriate value."/> <param name="cutoff2" type="text" value="12" label="Cutoff 2" help="Offset 2, which is used for P-sites calculation. Default is '12' but it varies a lot in different datasets. Please run 'ribotaper part 2' to deterimine a appropriate value."/> <param name="cutoff3" type="text" value="12" label="Cutoff 3" help="Offset 3, which is used for P-sites calculation. Default is '12' but it varies a lot in different datasets. Please run 'ribotaper part 2' to deterimine a appropriate value."/> </inputs> <outputs> <data name="output1" type="data" format="pdf" from_work_dir="quality_check_plots.pdf" label="QC plots"/> <data name="output2" type="data" format="tabular" from_work_dir="ORFs_genes_found" label="Summary of translated ORFs"/> <data name="output3" type="data" format="tabular" from_work_dir="ORFs_max" label="Translated ORFs (max)"/> <data name="output4" type="data" format="tabular" from_work_dir="ORFs_max_filt" label="Translated ORFs (max_filt)"/> <data name="output5" type="data" format="bed" from_work_dir="translated_ORFs_sorted.bed" label="Translated ORFs (sorted)"/> <data name="output6" type="data" format="bed" from_work_dir="translated_ORFs_filtered_sorted.bed" label="Translated ORFs (filtered/sorted)"/> <data name="output7" type="data" format="fasta" from_work_dir="protein_db_max.fasta" label="Protein DB"/> <data name="output8" type="data" format="pdf" from_work_dir="Final_ORF_results.pdf" label="ORF categories (length/coverage)"/> </outputs> <tests> <test> <param name="annotation_path" value="annotation_path.tgz" ftype="compressed_archive"/> <param name="ribo_bam" value="test_ribo.bam"/> <param name="rna_bam" value="test_rna.bam"/> <param name="read_lenghts_ribo1" value="26"/> <param name="read_lenghts_ribo2" value="28"/> <param name="read_lenghts_ribo3" value="29"/> <param name="cutoff1" value="9"/> <param name="cutoff2" value="12"/> <param name="cutoff3" value="12"/> <output name="output2" file="ORFs_genes_found"/> </test> </tests> <help><![CDATA[ RiboTaper is an analysis pipeline for Ribosome Profiling (Ribo-seq) experiments, which exploits the triplet periodicity of ribosomal footprints to call translated regions. See https://ohlerlab.mdc-berlin.de/software/RiboTaper_126/ for details. The Ribotaper Galaxy tool set consists of three tools: - ``ribotaper part 1``: creation of annotation files - ``ribotaper part 2``: metagene analysis for P-sites definition - ``ribotaper part 3``: ribosome profiling The order of execution should follow: ``ribotaper part 1, part 2 and part 3``. The current tool is ``ribotaper part 3``, ribosome profiling. Outputs -------- **QC plots**: This plot provides the user statistics about the Ribo-seq and RNA seq data used, together with the assessment of the P-sites calculations. Important values are the pie chart showing the agreement between the frame (defined by the P-sites position) and the annotated frame. Reliable P-sites calculations produce an agreement above 90%. Very important are also the length/coverage statistics for the Ribo-seq (bottom right): This shows how the P-site calculations can be used to detect active translation in regions of different length and coverage, in a way the user can estimate the precision of the Ribo-seq data, and understand the level or resolution the data allows. **Summary of translated ORFs**: Tab-separated values for the number of ORFs found and their corresponding genes, for the different ORF categories. **Translated ORFs (max, max_filt)**: Tab-separated file containing information about detected ORFs. Translated ORFs (max_filt) contains ORFs filtered for excessive multimapping and ORFs in non-coding genes overlapping known coding regions (recommended for further analysis). **Protein DB**: Fasta file of the detected ORFs peptide sequence, suitable as an alternative protein database (not filtered for multimapping) **Translated ORFs (sorted, filtered/sorted)**: BED files with genomic coordinates for the detected ORFs. The total number of P-sites along the ORF is reported on the 5th column. **ORF categories (length/coverage)**: PDF file containing info about the number of ORFs found, together with their length and coverage per category/annotation. Important notes ---------------- - We ran the RiboTaper analysis on an SGE cluster, using 7 cores and h_vmem 8G. For each dataset, the complete RiboTaper workflow (from the bam files to final results) took ~ 1 day. - The current RiboTaper framework is not designed to identify and quantify ORFs on different transcripts. This means the transcript annotation is crucial. - Be careful about using scaffolds, both in the genome and GTF files, which may slow the whole pipeline. ]]></help> <citations> <citation type="doi">10.1038/nmeth.3688</citation> </citations> </tool>