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1 This package contains a Galaxy workflow for the detection and incorporation of single amino acid polymorphism (SAP) into a custom proteomics search database.
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2
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3 The workflow aligns RNA-Seq reads to the organism's genome using Tophat, call SNPs using SAMtools mpileup command, and annotates the SNPs that reside within protein-coding regions using SNPeff.
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4 Galaxy tool "SNPeff to Peptide Fasta" generates peptide sequences with the polymorhisms.
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5
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6 See http://www.galaxyproject.org for information about the Galaxy Project.
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7
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8
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9 Availability
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10 ============
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11
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12 This workflow is available to download and/or install from the main
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13 Galaxy Tool Shed:
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14
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15 http://toolshed.g2.bx.psu.edu/view/galaxyp/proteomics_rnaseq_sap_db_workflow
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16
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17
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18 Reference Data
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19 ==============
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20
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21 For Human RNAseq data this workflow was tested with a genome build named "GRCh37_canon" using reference data from:
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22
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23 * ftp://ftp.ensembl.org/pub/release-73/fasta/homo_sapiens/dna/Homo_sapiens.GRCh37.73.dna.chromosome.[1-9XY]*.fa.gz
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24 * ftp://ftp.ensembl.org/pub/release-73/fasta/homo_sapiens/pep/Homo_sapiens.GRCh37.73.pep.all.fa.gz
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25 * ftp://ftp.ensembl.org/pub/release-73/gtf/homo_sapiens/Homo_sapiens.GRCh37.73.gtf.gz
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26
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27
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28 For Mouse RNAseq data this workflow was tested with a genome build named "GRCm38_canon" using reference data from:
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29
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30 * ftp://ftp.ensembl.org/pub/release-73/fasta/mus_musculus/dna/Mus_musculus.GRCm38.73.dna.chromosome.[1-9XY]*.fa.gz
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31 * ftp://ftp.ensembl.org/pub/release-73/fasta/mus_musculus/pep/Mus_musculus.GRCm38.73.pep.all.fa.gz
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32 * ftp://ftp.ensembl.org/pub/release-73/gtf/mus_musculus/Mus_musculus.GRCm38.73.gtf.gz
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33
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34
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35 Genome builds "GRCh37_canon" and "GRCm38_canon" were built according to instructions in: https://wiki.galaxyproject.org/Admin/NGS%20Local%20Setup
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36 The builds used only the standard chromosomesi sequences from the reference fasta; the other sequences were filtered out.
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37 The GTF was filtered to retain only those entries that referenced the standard chromosomes.
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38
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39
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40 Dependencies
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41 ============
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42
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43 These dependencies should be resolved automatically via the Galaxy Tool Shed:
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44
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45 * http://toolshed.g2.bx.psu.edu/view/devteam/tophat
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46 * http://toolshed.g2.bx.psu.edu/view/devteam/samtools_mpileup
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47 * http://toolshed.g2.bx.psu.edu/view/nilesh/bcftools
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48 * http://toolshed.g2.bx.psu.edu/view/iuc/snpeff
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49 * http://toolshed.g2.bx.psu.edu/view/jjohnson/snpeff_to_peptides
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50
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51 History
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52 =======
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53
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54 ======= ======================================================================
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55 Version Changes
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56 ------- ----------------------------------------------------------------------
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57 v0.0.1 - Initial release to Tool Shed (March, 2014)
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58 ======= ======================================================================
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59
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60
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