Mercurial > repos > pjbriggs > amplicon_analysis_pipeline
annotate README.rst @ 10:b47cf1d6fb47 draft
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| author | pjbriggs | 
|---|---|
| date | Mon, 18 Jun 2018 07:21:57 -0400 | 
| parents | bbfc9638ba84 | 
| children | b5e43dfe07ac | 
| rev | line source | 
|---|---|
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0
 
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit ba3e5b591407db52a586361efb21927c8171ec0e
 
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1 Amplicon_analysis-galaxy | 
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2 ======================== | 
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3 | 
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4 A Galaxy tool wrapper to Mauro Tutino's ``Amplicon_analysis`` pipeline | 
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5 script at https://github.com/MTutino/Amplicon_analysis | 
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6 | 
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7 The pipeline can analyse paired-end 16S rRNA data from Illumina Miseq | 
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8 (Casava >= 1.8) and performs the following operations: | 
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9 | 
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10 * QC and clean up of input data | 
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11 * Removal of singletons and chimeras and building of OTU table | 
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12 and phylogenetic tree | 
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13 * Beta and alpha diversity of analysis | 
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14 | 
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15 Usage documentation | 
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16 =================== | 
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17 | 
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18 Usage of the tool (including required inputs) is documented within | 
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19 the ``help`` section of the tool XML. | 
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20 | 
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21 Installing the tool in a Galaxy instance | 
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22 ======================================== | 
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23 | 
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1
 
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24 The following sections describe how to install the tool files, | 
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25 dependencies and reference data, and how to configure the Galaxy | 
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26 instance to detect the dependencies and reference data correctly | 
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27 at run time. | 
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28 | 
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29 1. Install the dependencies | 
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30 --------------------------- | 
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31 | 
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32 The ``install_tool_deps.sh`` script can be used to fetch and install the | 
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33 dependencies locally, for example:: | 
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34 | 
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35 install_tool_deps.sh /path/to/local_tool_dependencies | 
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36 | 
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37 This can take some time to complete. When finished it should have | 
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38 created a set of directories containing the dependencies under the | 
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39 specified top level directory. | 
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40 | 
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41 2. Install the tool files | 
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42 ------------------------- | 
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43 | 
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1
 
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44 The core tool is hosted on the Galaxy toolshed, so it can be installed | 
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45 directly from there (this is the recommended route): | 
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46 | 
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47 * https://toolshed.g2.bx.psu.edu/view/pjbriggs/amplicon_analysis_pipeline/ | 
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48 | 
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49 Alternatively it can be installed manually; in this case there are two | 
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50 files to install: | 
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51 | 
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52 * ``amplicon_analysis_pipeline.xml`` (the Galaxy tool definition) | 
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53 * ``amplicon_analysis_pipeline.py`` (the Python wrapper script) | 
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54 | 
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55 Put these in a directory that is visible to Galaxy (e.g. a | 
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56 ``tools/Amplicon_analysis/`` folder), and modify the ``tools_conf.xml`` | 
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57 file to tell Galaxy to offer the tool by adding the line e.g.:: | 
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58 | 
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59 <tool file="Amplicon_analysis/amplicon_analysis_pipeline.xml" /> | 
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60 | 
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61 3. Install the reference data | 
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62 ----------------------------- | 
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63 | 
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64 The script ``References.sh`` from the pipeline package at | 
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65 https://github.com/MTutino/Amplicon_analysis can be run to install | 
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66 the reference data, for example:: | 
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67 | 
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68 cd /path/to/pipeline/data | 
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69 wget https://github.com/MTutino/Amplicon_analysis/raw/master/References.sh | 
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70 /bin/bash ./References.sh | 
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71 | 
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72 will install the data in ``/path/to/pipeline/data``. | 
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73 | 
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74 **NB** The final amount of data downloaded and uncompressed will be | 
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75 around 6GB. | 
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76 | 
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77 4. Configure dependencies and reference data in Galaxy | 
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78 ------------------------------------------------------ | 
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79 | 
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80 The final steps are to make your Galaxy installation aware of the | 
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81 tool dependencies and reference data, so it can locate them both when | 
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 | 
82 the tool is run. | 
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83 | 
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84 To target the tool dependencies installed previously, add the | 
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85 following lines to the ``dependency_resolvers_conf.xml`` file in the | 
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86 Galaxy ``config`` directory:: | 
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87 | 
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88 <dependency_resolvers> | 
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89 ... | 
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90 <galaxy_packages base_path="/path/to/local_tool_dependencies" /> | 
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91 <galaxy_packages base_path="/path/to/local_tool_dependencies" versionless="true" /> | 
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92 ... | 
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93 </dependency_resolvers> | 
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94 | 
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95 (NB it is recommended to place these *before* the ``<conda ... />`` | 
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96 resolvers) | 
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97 | 
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98 (If you're not familiar with dependency resolvers in Galaxy then | 
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99 see the documentation at | 
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100 https://docs.galaxyproject.org/en/master/admin/dependency_resolvers.html | 
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101 for more details.) | 
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102 | 
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103 The tool locates the reference data via an environment variable called | 
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104 ``AMPLICON_ANALYSIS_REF_DATA_PATH``, which needs to set to the parent | 
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105 directory where the reference data has been installed. | 
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106 | 
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107 There are various ways to do this, depending on how your Galaxy | 
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108 installation is configured: | 
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109 | 
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110 * **For local instances:** add a line to set it in the | 
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111 ``config/local_env.sh`` file of your Galaxy installation, e.g.:: | 
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112 | 
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113 export AMPLICON_ANALYSIS_REF_DATA_PATH=/path/to/pipeline/data | 
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114 | 
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115 * **For production instances:** set the value in the ``job_conf.xml`` | 
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116 configuration file, e.g.:: | 
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117 | 
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118 <destination id="amplicon_analysis"> | 
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119 <env id="AMPLICON_ANALYSIS_REF_DATA_PATH">/path/to/pipeline/data</env> | 
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120 </destination> | 
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121 | 
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122 and then specify that the pipeline tool uses this destination:: | 
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123 | 
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124 <tool id="amplicon_analysis_pipeline" destination="amplicon_analysis"/> | 
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125 | 
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126 (For more about job destinations see the Galaxy documentation at | 
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127 https://galaxyproject.org/admin/config/jobs/#job-destinations) | 
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128 | 
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129 5. Enable rendering of HTML outputs from pipeline | 
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130 ------------------------------------------------- | 
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131 | 
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132 To ensure that HTML outputs are displayed correctly in Galaxy | 
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133 (for example the Vsearch OTU table heatmaps), Galaxy needs to be | 
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134 configured not to sanitize the outputs from the ``Amplicon_analysis`` | 
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135 tool. | 
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136 | 
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137 Either: | 
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138 | 
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139 * **For local instances:** set ``sanitize_all_html = False`` in | 
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140 ``config/galaxy.ini`` (nb don't do this on production servers or | 
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141 public instances!); or | 
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142 | 
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143 * **For production instances:** add the ``Amplicon_analysis`` tool | 
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144 to the display whitelist in the Galaxy instance: | 
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145 | 
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146 - Set ``sanitize_whitelist_file = config/whitelist.txt`` in | 
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147 ``config/galaxy.ini`` and restart Galaxy; | 
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148 - Go to ``Admin>Manage Display Whitelist``, check the box for | 
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149 ``Amplicon_analysis`` (hint: use your browser's 'find-in-page' | 
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150 search function to help locate it) and click on | 
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151 ``Submit new whitelist`` to update the settings. | 
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152 | 
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153 Additional details | 
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154 ================== | 
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155 | 
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156 Some other things to be aware of: | 
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157 | 
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158 * Note that using the Silva database requires a minimum of 18Gb RAM | 
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159 | 
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160 Known problems | 
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161 ============== | 
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162 | 
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163 * Only the ``VSEARCH`` pipeline in Mauro's script is currently | 
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164 available via the Galaxy tool; the ``USEARCH`` and ``QIIME`` | 
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165 pipelines have yet to be implemented. | 
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166 * The images in the tool help section are not visible if the | 
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167 tool has been installed locally, or if it has been installed in | 
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168 a Galaxy instance which is served from a subdirectory. | 
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169 | 
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170 These are both problems with Galaxy and not the tool, see | 
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171 https://github.com/galaxyproject/galaxy/issues/4490 and | 
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172 https://github.com/galaxyproject/galaxy/issues/1676 | 
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173 | 
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174 Appendix: availability of tool dependencies | 
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175 =========================================== | 
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176 | 
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177 The tool takes its dependencies from the underlying pipeline script (see | 
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178 https://github.com/MTutino/Amplicon_analysis/blob/master/README.md | 
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179 for details). | 
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180 | 
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181 As noted above, currently the ``install_tool_deps.sh`` script can be | 
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182 used to manually install the dependencies for a local tool install. | 
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183 | 
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184 In principle these should also be available if the tool were installed | 
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185 from a toolshed. However it would be preferrable in this case to get as | 
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186 many of the dependencies as possible via the ``conda`` dependency | 
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187 resolver. | 
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188 | 
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189 The following are known to be available via conda, with the required | 
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190 version: | 
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191 | 
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192 - cutadapt 1.8.1 | 
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193 - sickle-trim 1.33 | 
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194 - bioawk 1.0 | 
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195 - fastqc 0.11.3 | 
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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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203 | 
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204 The following dependencies are currently unavailable: | 
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205 | 
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206 - fasta_number (need 02jun2015) | 
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207 - microbiomeutil (need r20110519) | 
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208 | 
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209 (NB usearch 6.1.544 and 8.0.1623 are special cases which must be | 
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210 handled outside of Galaxy's dependency management systems.) | 
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211 | 
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212 History | 
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213 ======= | 
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214 | 
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215 ========== ====================================================================== | 
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216 Version Changes | 
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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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224 1.1.0 First official version on Galaxy toolshed. | 
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225 1.0.6 Expand inline documentation to provide detailed usage guidance. | 
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226 1.0.5 Updates including: | 
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227 | 
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228 - Capture read counts from quality control as new output dataset | 
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229 - Capture FastQC per-base quality boxplots for each sample as | 
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230 new output dataset | 
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231 - Add support for -l option (sliding window length for trimming) | 
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232 - Default for -L set to "200" | 
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233 1.0.4 Various updates: | 
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234 | 
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235 - Additional outputs are captured when a "Categories" file is | 
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236 supplied (alpha diversity rarefaction curves and boxplots) | 
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237 - Sample names derived from Fastqs in a collection of pairs | 
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238 are trimmed to SAMPLE_S* (for Illumina-style Fastq filenames) | 
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239 - Input Fastqs can now be of more general ``fastq`` type | 
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240 - Log file outputs are captured in new output dataset | 
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241 - User can specify a "title" for the job which is copied into | 
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242 the dataset names (to distinguish outputs from different runs) | 
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243 - Improved detection and reporting of problems with input | 
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244 Metatable | 
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245 1.0.3 Take the sample names from the collection dataset names when | 
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246 using collection as input (this is now the default input mode); | 
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247 collect additional output dataset; disable ``usearch``-based | 
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248 pipelines (i.e. ``UPARSE`` and ``QIIME``). | 
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249 1.0.2 Enable support for FASTQs supplied via dataset collections and | 
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250 fix some broken output datasets. | 
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251 1.0.1 Initial version | 
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252 ========== ====================================================================== | 
