Mercurial > repos > pjbriggs > amplicon_analysis_pipeline
annotate README.rst @ 13:c87b166cbfe1 draft
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| author | pjbriggs | 
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| date | Mon, 18 Jun 2018 10:05:58 -0400 | 
| parents | bbfc9638ba84 | 
| children | b5e43dfe07ac | 
| rev | line source | 
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit ba3e5b591407db52a586361efb21927c8171ec0e
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changeset | 1 Amplicon_analysis-galaxy | 
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changeset | 2 ======================== | 
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changeset | 3 | 
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changeset | 4 A Galaxy tool wrapper to Mauro Tutino's ``Amplicon_analysis`` pipeline | 
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changeset | 5 script at https://github.com/MTutino/Amplicon_analysis | 
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changeset | 6 | 
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changeset | 7 The pipeline can analyse paired-end 16S rRNA data from Illumina Miseq | 
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changeset | 8 (Casava >= 1.8) and performs the following operations: | 
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changeset | 9 | 
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changeset | 10 * QC and clean up of input data | 
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changeset | 11 * Removal of singletons and chimeras and building of OTU table | 
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changeset | 12 and phylogenetic tree | 
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changeset | 13 * Beta and alpha diversity of analysis | 
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changeset | 14 | 
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changeset | 15 Usage documentation | 
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changeset | 16 =================== | 
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changeset | 17 | 
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changeset | 18 Usage of the tool (including required inputs) is documented within | 
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changeset | 19 the ``help`` section of the tool XML. | 
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changeset | 20 | 
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changeset | 21 Installing the tool in a Galaxy instance | 
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changeset | 22 ======================================== | 
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changeset | 23 | 
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changeset | 24 The following sections describe how to install the tool files, | 
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changeset | 25 dependencies and reference data, and how to configure the Galaxy | 
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changeset | 26 instance to detect the dependencies and reference data correctly | 
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changeset | 27 at run time. | 
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changeset | 28 | 
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changeset | 29 1. Install the dependencies | 
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changeset | 30 --------------------------- | 
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changeset | 31 | 
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changeset | 32 The ``install_tool_deps.sh`` script can be used to fetch and install the | 
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changeset | 33 dependencies locally, for example:: | 
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changeset | 34 | 
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changeset | 35 install_tool_deps.sh /path/to/local_tool_dependencies | 
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changeset | 36 | 
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changeset | 37 This can take some time to complete. When finished it should have | 
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changeset | 38 created a set of directories containing the dependencies under the | 
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changeset | 39 specified top level directory. | 
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changeset | 40 | 
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changeset | 41 2. Install the tool files | 
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changeset | 42 ------------------------- | 
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changeset | 43 | 
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changeset | 44 The core tool is hosted on the Galaxy toolshed, so it can be installed | 
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changeset | 45 directly from there (this is the recommended route): | 
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changeset | 46 | 
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changeset | 47 * https://toolshed.g2.bx.psu.edu/view/pjbriggs/amplicon_analysis_pipeline/ | 
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changeset | 48 | 
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changeset | 49 Alternatively it can be installed manually; in this case there are two | 
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changeset | 50 files to install: | 
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changeset | 51 | 
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changeset | 52 * ``amplicon_analysis_pipeline.xml`` (the Galaxy tool definition) | 
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changeset | 53 * ``amplicon_analysis_pipeline.py`` (the Python wrapper script) | 
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changeset | 54 | 
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changeset | 55 Put these in a directory that is visible to Galaxy (e.g. a | 
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changeset | 56 ``tools/Amplicon_analysis/`` folder), and modify the ``tools_conf.xml`` | 
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changeset | 57 file to tell Galaxy to offer the tool by adding the line e.g.:: | 
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changeset | 58 | 
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changeset | 59 <tool file="Amplicon_analysis/amplicon_analysis_pipeline.xml" /> | 
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changeset | 60 | 
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changeset | 61 3. Install the reference data | 
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changeset | 62 ----------------------------- | 
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changeset | 63 | 
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changeset | 64 The script ``References.sh`` from the pipeline package at | 
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changeset | 65 https://github.com/MTutino/Amplicon_analysis can be run to install | 
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changeset | 66 the reference data, for example:: | 
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changeset | 67 | 
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changeset | 68 cd /path/to/pipeline/data | 
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changeset | 69 wget https://github.com/MTutino/Amplicon_analysis/raw/master/References.sh | 
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changeset | 70 /bin/bash ./References.sh | 
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changeset | 71 | 
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changeset | 72 will install the data in ``/path/to/pipeline/data``. | 
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changeset | 73 | 
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changeset | 74 **NB** The final amount of data downloaded and uncompressed will be | 
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changeset | 75 around 6GB. | 
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changeset | 76 | 
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changeset | 77 4. Configure dependencies and reference data in Galaxy | 
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changeset | 78 ------------------------------------------------------ | 
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changeset | 79 | 
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changeset | 80 The final steps are to make your Galaxy installation aware of the | 
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changeset | 81 tool dependencies and reference data, so it can locate them both when | 
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changeset | 82 the tool is run. | 
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changeset | 83 | 
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changeset | 84 To target the tool dependencies installed previously, add the | 
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changeset | 85 following lines to the ``dependency_resolvers_conf.xml`` file in the | 
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changeset | 86 Galaxy ``config`` directory:: | 
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changeset | 87 | 
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changeset | 88 <dependency_resolvers> | 
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changeset | 89 ... | 
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changeset | 90 <galaxy_packages base_path="/path/to/local_tool_dependencies" /> | 
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changeset | 91 <galaxy_packages base_path="/path/to/local_tool_dependencies" versionless="true" /> | 
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changeset | 92 ... | 
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changeset | 93 </dependency_resolvers> | 
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changeset | 94 | 
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changeset | 95 (NB it is recommended to place these *before* the ``<conda ... />`` | 
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changeset | 96 resolvers) | 
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changeset | 97 | 
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changeset | 98 (If you're not familiar with dependency resolvers in Galaxy then | 
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changeset | 99 see the documentation at | 
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changeset | 100 https://docs.galaxyproject.org/en/master/admin/dependency_resolvers.html | 
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changeset | 101 for more details.) | 
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changeset | 102 | 
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changeset | 103 The tool locates the reference data via an environment variable called | 
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changeset | 104 ``AMPLICON_ANALYSIS_REF_DATA_PATH``, which needs to set to the parent | 
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changeset | 105 directory where the reference data has been installed. | 
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changeset | 106 | 
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changeset | 107 There are various ways to do this, depending on how your Galaxy | 
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changeset | 108 installation is configured: | 
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changeset | 109 | 
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changeset | 110 * **For local instances:** add a line to set it in the | 
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changeset | 111 ``config/local_env.sh`` file of your Galaxy installation, e.g.:: | 
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changeset | 112 | 
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changeset | 113 export AMPLICON_ANALYSIS_REF_DATA_PATH=/path/to/pipeline/data | 
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changeset | 114 | 
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changeset | 115 * **For production instances:** set the value in the ``job_conf.xml`` | 
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changeset | 116 configuration file, e.g.:: | 
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changeset | 117 | 
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changeset | 118 <destination id="amplicon_analysis"> | 
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changeset | 119 <env id="AMPLICON_ANALYSIS_REF_DATA_PATH">/path/to/pipeline/data</env> | 
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changeset | 120 </destination> | 
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changeset | 121 | 
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changeset | 122 and then specify that the pipeline tool uses this destination:: | 
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changeset | 123 | 
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changeset | 124 <tool id="amplicon_analysis_pipeline" destination="amplicon_analysis"/> | 
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changeset | 125 | 
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changeset | 126 (For more about job destinations see the Galaxy documentation at | 
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changeset | 127 https://galaxyproject.org/admin/config/jobs/#job-destinations) | 
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changeset | 128 | 
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changeset | 129 5. Enable rendering of HTML outputs from pipeline | 
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changeset | 130 ------------------------------------------------- | 
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changeset | 131 | 
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changeset | 132 To ensure that HTML outputs are displayed correctly in Galaxy | 
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changeset | 133 (for example the Vsearch OTU table heatmaps), Galaxy needs to be | 
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changeset | 134 configured not to sanitize the outputs from the ``Amplicon_analysis`` | 
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changeset | 135 tool. | 
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changeset | 136 | 
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changeset | 137 Either: | 
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changeset | 138 | 
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changeset | 139 * **For local instances:** set ``sanitize_all_html = False`` in | 
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changeset | 140 ``config/galaxy.ini`` (nb don't do this on production servers or | 
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changeset | 141 public instances!); or | 
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changeset | 142 | 
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changeset | 143 * **For production instances:** add the ``Amplicon_analysis`` tool | 
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changeset | 144 to the display whitelist in the Galaxy instance: | 
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changeset | 145 | 
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changeset | 146 - Set ``sanitize_whitelist_file = config/whitelist.txt`` in | 
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changeset | 147 ``config/galaxy.ini`` and restart Galaxy; | 
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changeset | 148 - Go to ``Admin>Manage Display Whitelist``, check the box for | 
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changeset | 149 ``Amplicon_analysis`` (hint: use your browser's 'find-in-page' | 
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changeset | 150 search function to help locate it) and click on | 
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changeset | 151 ``Submit new whitelist`` to update the settings. | 
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changeset | 152 | 
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changeset | 153 Additional details | 
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changeset | 154 ================== | 
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changeset | 155 | 
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changeset | 156 Some other things to be aware of: | 
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changeset | 157 | 
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changeset | 158 * Note that using the Silva database requires a minimum of 18Gb RAM | 
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changeset | 159 | 
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changeset | 160 Known problems | 
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changeset | 161 ============== | 
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changeset | 162 | 
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changeset | 163 * Only the ``VSEARCH`` pipeline in Mauro's script is currently | 
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changeset | 164 available via the Galaxy tool; the ``USEARCH`` and ``QIIME`` | 
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changeset | 165 pipelines have yet to be implemented. | 
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changeset | 166 * The images in the tool help section are not visible if the | 
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changeset | 167 tool has been installed locally, or if it has been installed in | 
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changeset | 168 a Galaxy instance which is served from a subdirectory. | 
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changeset | 169 | 
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changeset | 170 These are both problems with Galaxy and not the tool, see | 
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changeset | 171 https://github.com/galaxyproject/galaxy/issues/4490 and | 
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changeset | 172 https://github.com/galaxyproject/galaxy/issues/1676 | 
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changeset | 173 | 
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changeset | 174 Appendix: availability of tool dependencies | 
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changeset | 175 =========================================== | 
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changeset | 176 | 
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changeset | 177 The tool takes its dependencies from the underlying pipeline script (see | 
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changeset | 178 https://github.com/MTutino/Amplicon_analysis/blob/master/README.md | 
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changeset | 179 for details). | 
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changeset | 180 | 
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changeset | 181 As noted above, currently the ``install_tool_deps.sh`` script can be | 
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changeset | 182 used to manually install the dependencies for a local tool install. | 
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changeset | 183 | 
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changeset | 184 In principle these should also be available if the tool were installed | 
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changeset | 185 from a toolshed. However it would be preferrable in this case to get as | 
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changeset | 186 many of the dependencies as possible via the ``conda`` dependency | 
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changeset | 187 resolver. | 
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changeset | 188 | 
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changeset | 189 The following are known to be available via conda, with the required | 
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changeset | 190 version: | 
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changeset | 191 | 
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changeset | 192 - cutadapt 1.8.1 | 
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changeset | 193 - sickle-trim 1.33 | 
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changeset | 194 - bioawk 1.0 | 
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changeset | 195 - fastqc 0.11.3 | 
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changeset | 196 - R 3.2.0 | 
| 5 | 197 - spades 3.5.0 | 
| 198 - qiime 1.8.0 | |
| 199 - blast-legacy 2.2.26 | |
| 200 - vsearch 1.1.3 | |
| 201 - fasta-splitter 0.2.4 | |
| 202 - rdp_classifier 2.2 | |
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changeset | 203 | 
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changeset | 204 The following dependencies are currently unavailable: | 
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changeset | 205 | 
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changeset | 206 - fasta_number (need 02jun2015) | 
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changeset | 207 - microbiomeutil (need r20110519) | 
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changeset | 208 | 
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changeset | 209 (NB usearch 6.1.544 and 8.0.1623 are special cases which must be | 
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changeset | 210 handled outside of Galaxy's dependency management systems.) | 
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changeset | 211 | 
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changeset | 212 History | 
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changeset | 213 ======= | 
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changeset | 214 | 
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changeset | 215 ========== ====================================================================== | 
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changeset | 216 Version Changes | 
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changeset | 217 ---------- ---------------------------------------------------------------------- | 
| 5 | 218 1.2.2.1 Update to get dependencies from bioconda | 
| 4 | 219 1.2.2.0 Updated to Amplicon_Analysis_Pipeline version 1.2.2 (removes | 
| 220 jackknifed analysis which is not captured by Galaxy tool) | |
| 2 | 221 1.2.1.0 Updated to Amplicon_Analysis_Pipeline version 1.2.1 (adds | 
| 222 option to use the Human Oral Microbiome Database v15.1, and | |
| 223 updates SILVA database to v123) | |
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changeset | 224 1.1.0 First official version on Galaxy toolshed. | 
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changeset | 225 1.0.6 Expand inline documentation to provide detailed usage guidance. | 
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changeset | 226 1.0.5 Updates including: | 
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changeset | 227 | 
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changeset | 228 - Capture read counts from quality control as new output dataset | 
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changeset | 229 - Capture FastQC per-base quality boxplots for each sample as | 
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changeset | 230 new output dataset | 
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changeset | 231 - Add support for -l option (sliding window length for trimming) | 
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changeset | 232 - Default for -L set to "200" | 
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changeset | 233 1.0.4 Various updates: | 
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changeset | 234 | 
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changeset | 235 - Additional outputs are captured when a "Categories" file is | 
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changeset | 236 supplied (alpha diversity rarefaction curves and boxplots) | 
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changeset | 237 - Sample names derived from Fastqs in a collection of pairs | 
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changeset | 238 are trimmed to SAMPLE_S* (for Illumina-style Fastq filenames) | 
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changeset | 239 - Input Fastqs can now be of more general ``fastq`` type | 
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changeset | 240 - Log file outputs are captured in new output dataset | 
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changeset | 241 - User can specify a "title" for the job which is copied into | 
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changeset | 242 the dataset names (to distinguish outputs from different runs) | 
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changeset | 243 - Improved detection and reporting of problems with input | 
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changeset | 244 Metatable | 
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changeset | 245 1.0.3 Take the sample names from the collection dataset names when | 
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changeset | 246 using collection as input (this is now the default input mode); | 
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changeset | 247 collect additional output dataset; disable ``usearch``-based | 
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changeset | 248 pipelines (i.e. ``UPARSE`` and ``QIIME``). | 
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changeset | 249 1.0.2 Enable support for FASTQs supplied via dataset collections and | 
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changeset | 250 fix some broken output datasets. | 
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changeset | 251 1.0.1 Initial version | 
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changeset | 252 ========== ====================================================================== | 
