Mercurial > repos > pjbriggs > amplicon_analysis_pipeline
annotate README.rst @ 32:d94f2d7e8574 draft
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| author | pjbriggs | 
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| date | Tue, 02 Oct 2018 10:18:02 -0400 | 
| parents | 18e6427c5e4a | 
| children | d19fca1c009c | 
| rev | line source | 
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0
 
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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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27
 
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32 If the tool is installed from the Galaxy toolshed (recommended) then | 
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33 the dependencies should be installed automatically and this step can | 
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34 be skipped. | 
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35 | 
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36 Otherwise the ``install_tool_deps.sh`` script can be used to fetch and | 
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37 install the dependencies locally, for example:: | 
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38 | 
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39 install_tool_deps.sh /path/to/local_tool_dependencies | 
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40 | 
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41 This can take some time to complete. When finished it should have | 
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42 created a set of directories containing the dependencies under the | 
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43 specified top level directory. | 
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44 | 
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27
 
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45 *NB ``install_tool_deps.sh`` is no longer being maintained* | 
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46 | 
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47 2. Install the tool files | 
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48 ------------------------- | 
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49 | 
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1
 
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50 The core tool is hosted on the Galaxy toolshed, so it can be installed | 
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51 directly from there (this is the recommended route): | 
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52 | 
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53 * https://toolshed.g2.bx.psu.edu/view/pjbriggs/amplicon_analysis_pipeline/ | 
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54 | 
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55 Alternatively it can be installed manually; in this case there are two | 
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56 files to install: | 
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57 | 
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58 * ``amplicon_analysis_pipeline.xml`` (the Galaxy tool definition) | 
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59 * ``amplicon_analysis_pipeline.py`` (the Python wrapper script) | 
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60 | 
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61 Put these in a directory that is visible to Galaxy (e.g. a | 
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62 ``tools/Amplicon_analysis/`` folder), and modify the ``tools_conf.xml`` | 
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63 file to tell Galaxy to offer the tool by adding the line e.g.:: | 
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64 | 
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65 <tool file="Amplicon_analysis/amplicon_analysis_pipeline.xml" /> | 
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66 | 
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67 3. Install the reference data | 
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68 ----------------------------- | 
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69 | 
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70 The script ``References.sh`` from the pipeline package at | 
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71 https://github.com/MTutino/Amplicon_analysis can be run to install | 
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72 the reference data, for example:: | 
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73 | 
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74 cd /path/to/pipeline/data | 
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75 wget https://github.com/MTutino/Amplicon_analysis/raw/master/References.sh | 
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76 /bin/bash ./References.sh | 
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77 | 
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78 will install the data in ``/path/to/pipeline/data``. | 
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79 | 
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80 **NB** The final amount of data downloaded and uncompressed will be | 
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81 around 6GB. | 
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82 | 
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83 4. Configure dependencies and reference data in Galaxy | 
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84 ------------------------------------------------------ | 
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85 | 
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86 The final steps are to make your Galaxy installation aware of the | 
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87 tool dependencies and reference data, so it can locate them both when | 
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88 the tool is run. | 
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89 | 
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90 To target the tool dependencies installed previously, add the | 
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91 following lines to the ``dependency_resolvers_conf.xml`` file in the | 
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92 Galaxy ``config`` directory:: | 
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93 | 
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94 <dependency_resolvers> | 
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95 ... | 
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96 <galaxy_packages base_path="/path/to/local_tool_dependencies" /> | 
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97 <galaxy_packages base_path="/path/to/local_tool_dependencies" versionless="true" /> | 
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98 ... | 
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99 </dependency_resolvers> | 
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100 | 
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101 (NB it is recommended to place these *before* the ``<conda ... />`` | 
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102 resolvers) | 
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103 | 
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104 (If you're not familiar with dependency resolvers in Galaxy then | 
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105 see the documentation at | 
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106 https://docs.galaxyproject.org/en/master/admin/dependency_resolvers.html | 
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107 for more details.) | 
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108 | 
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109 The tool locates the reference data via an environment variable called | 
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110 ``AMPLICON_ANALYSIS_REF_DATA_PATH``, which needs to set to the parent | 
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111 directory where the reference data has been installed. | 
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112 | 
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113 There are various ways to do this, depending on how your Galaxy | 
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114 installation is configured: | 
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115 | 
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116 * **For local instances:** add a line to set it in the | 
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117 ``config/local_env.sh`` file of your Galaxy installation, e.g.:: | 
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118 | 
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119 export AMPLICON_ANALYSIS_REF_DATA_PATH=/path/to/pipeline/data | 
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120 | 
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121 * **For production instances:** set the value in the ``job_conf.xml`` | 
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122 configuration file, e.g.:: | 
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123 | 
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124 <destination id="amplicon_analysis"> | 
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125 <env id="AMPLICON_ANALYSIS_REF_DATA_PATH">/path/to/pipeline/data</env> | 
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126 </destination> | 
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127 | 
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128 and then specify that the pipeline tool uses this destination:: | 
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129 | 
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130 <tool id="amplicon_analysis_pipeline" destination="amplicon_analysis"/> | 
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131 | 
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132 (For more about job destinations see the Galaxy documentation at | 
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133 https://galaxyproject.org/admin/config/jobs/#job-destinations) | 
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134 | 
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135 5. Enable rendering of HTML outputs from pipeline | 
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136 ------------------------------------------------- | 
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137 | 
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138 To ensure that HTML outputs are displayed correctly in Galaxy | 
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139 (for example the Vsearch OTU table heatmaps), Galaxy needs to be | 
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140 configured not to sanitize the outputs from the ``Amplicon_analysis`` | 
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141 tool. | 
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142 | 
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143 Either: | 
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144 | 
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145 * **For local instances:** set ``sanitize_all_html = False`` in | 
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146 ``config/galaxy.ini`` (nb don't do this on production servers or | 
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147 public instances!); or | 
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148 | 
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149 * **For production instances:** add the ``Amplicon_analysis`` tool | 
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150 to the display whitelist in the Galaxy instance: | 
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151 | 
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152 - Set ``sanitize_whitelist_file = config/whitelist.txt`` in | 
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153 ``config/galaxy.ini`` and restart Galaxy; | 
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154 - Go to ``Admin>Manage Display Whitelist``, check the box for | 
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155 ``Amplicon_analysis`` (hint: use your browser's 'find-in-page' | 
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156 search function to help locate it) and click on | 
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157 ``Submit new whitelist`` to update the settings. | 
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158 | 
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159 Additional details | 
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160 ================== | 
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161 | 
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162 Some other things to be aware of: | 
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163 | 
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164 * Note that using the Silva database requires a minimum of 18Gb RAM | 
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165 | 
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166 Known problems | 
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167 ============== | 
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168 | 
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169 * Only the ``VSEARCH`` pipeline in Mauro's script is currently | 
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170 available via the Galaxy tool; the ``USEARCH`` and ``QIIME`` | 
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171 pipelines have yet to be implemented. | 
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172 * The images in the tool help section are not visible if the | 
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173 tool has been installed locally, or if it has been installed in | 
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174 a Galaxy instance which is served from a subdirectory. | 
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175 | 
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176 These are both problems with Galaxy and not the tool, see | 
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177 https://github.com/galaxyproject/galaxy/issues/4490 and | 
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178 https://github.com/galaxyproject/galaxy/issues/1676 | 
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179 | 
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180 History | 
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181 ======= | 
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182 | 
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183 ========== ====================================================================== | 
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184 Version Changes | 
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185 ---------- ---------------------------------------------------------------------- | 
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27
 
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186 1.2.3.0 Updated to Amplicon_Analysis_Pipeline version 1.2.3; install | 
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30
 
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187 dependencies from bioconda and via tool_dependencies.xml. | 
| 4 | 188 1.2.2.0 Updated to Amplicon_Analysis_Pipeline version 1.2.2 (removes | 
| 189 jackknifed analysis which is not captured by Galaxy tool) | |
| 2 | 190 1.2.1.0 Updated to Amplicon_Analysis_Pipeline version 1.2.1 (adds | 
| 191 option to use the Human Oral Microbiome Database v15.1, and | |
| 192 updates SILVA database to v123) | |
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0
 
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193 1.1.0 First official version on Galaxy toolshed. | 
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194 1.0.6 Expand inline documentation to provide detailed usage guidance. | 
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195 1.0.5 Updates including: | 
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196 | 
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197 - Capture read counts from quality control as new output dataset | 
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198 - Capture FastQC per-base quality boxplots for each sample as | 
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199 new output dataset | 
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200 - Add support for -l option (sliding window length for trimming) | 
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201 - Default for -L set to "200" | 
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202 1.0.4 Various updates: | 
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203 | 
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204 - Additional outputs are captured when a "Categories" file is | 
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205 supplied (alpha diversity rarefaction curves and boxplots) | 
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206 - Sample names derived from Fastqs in a collection of pairs | 
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207 are trimmed to SAMPLE_S* (for Illumina-style Fastq filenames) | 
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208 - Input Fastqs can now be of more general ``fastq`` type | 
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209 - Log file outputs are captured in new output dataset | 
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210 - User can specify a "title" for the job which is copied into | 
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211 the dataset names (to distinguish outputs from different runs) | 
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212 - Improved detection and reporting of problems with input | 
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213 Metatable | 
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214 1.0.3 Take the sample names from the collection dataset names when | 
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215 using collection as input (this is now the default input mode); | 
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216 collect additional output dataset; disable ``usearch``-based | 
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217 pipelines (i.e. ``UPARSE`` and ``QIIME``). | 
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218 1.0.2 Enable support for FASTQs supplied via dataset collections and | 
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219 fix some broken output datasets. | 
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220 1.0.1 Initial version | 
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221 ========== ====================================================================== | 
