Mercurial > repos > pjbriggs > amplicon_analysis_pipeline
annotate README.rst @ 2:43d6f81bc667 draft
Updated to Amplicon_Analysis_pipeline v1.2.2.
author | pjbriggs |
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date | Wed, 13 Jun 2018 07:45:06 -0400 |
parents | 1c1902e12caf |
children | 3ab198df8f3f |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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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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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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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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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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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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30 --------------------------- |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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31 |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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32 The ``install_tool_deps.sh`` script can be used to fetch and install the |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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33 dependencies locally, for example:: |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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34 |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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35 install_tool_deps.sh /path/to/local_tool_dependencies |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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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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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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38 created a set of directories containing the dependencies under the |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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39 specified top level directory. |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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40 |
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41 2. Install the tool files |
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42 ------------------------- |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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43 |
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44 The core tool is hosted on the Galaxy toolshed, so it can be installed |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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45 directly from there (this is the recommended route): |
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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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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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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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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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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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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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planemo upload for repository https://github.com/pjbriggs/Amplicon_analysis-galaxy commit b63924933a03255872077beb4d0fde49d77afa92
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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 |
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197 |
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198 Some dependencies are available but with the "wrong" versions: |
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199 |
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200 - spades (need 3.5.0) |
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201 - qiime (need 1.8.0) |
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202 - blast (need 2.2.26) |
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203 - vsearch (need 1.1.3) |
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204 |
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205 The following dependencies are currently unavailable: |
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206 |
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207 - fasta_number (need 02jun2015) |
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208 - fasta-splitter (need 0.2.4) |
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209 - rdp_classifier (need 2.2) |
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210 - microbiomeutil (need r20110519) |
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211 |
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212 (NB usearch 6.1.544 and 8.0.1623 are special cases which must be |
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213 handled outside of Galaxy's dependency management systems.) |
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214 |
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215 History |
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216 ======= |
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217 |
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218 ========== ====================================================================== |
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219 Version Changes |
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220 ---------- ---------------------------------------------------------------------- |
2 | 221 1.2.2.0 Updated to Amplicon_Analysis_Pipeline version 1.2.2 (removes |
222 jackknifed analysis which is not captured by Galaxy tool) | |
1 | 223 1.2.1.0 Updated to Amplicon_Analysis_Pipeline version 1.2.1 (adds |
224 option to use the Human Oral Microbiome Database v15.1, and | |
225 updates SILVA database to v123) | |
0
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226 1.1.0 First official version on Galaxy toolshed. |
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227 1.0.6 Expand inline documentation to provide detailed usage guidance. |
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228 1.0.5 Updates including: |
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229 |
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230 - Capture read counts from quality control as new output dataset |
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231 - Capture FastQC per-base quality boxplots for each sample as |
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232 new output dataset |
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233 - Add support for -l option (sliding window length for trimming) |
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234 - Default for -L set to "200" |
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235 1.0.4 Various updates: |
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236 |
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237 - Additional outputs are captured when a "Categories" file is |
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238 supplied (alpha diversity rarefaction curves and boxplots) |
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239 - Sample names derived from Fastqs in a collection of pairs |
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240 are trimmed to SAMPLE_S* (for Illumina-style Fastq filenames) |
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241 - Input Fastqs can now be of more general ``fastq`` type |
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242 - Log file outputs are captured in new output dataset |
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243 - User can specify a "title" for the job which is copied into |
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244 the dataset names (to distinguish outputs from different runs) |
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245 - Improved detection and reporting of problems with input |
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246 Metatable |
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247 1.0.3 Take the sample names from the collection dataset names when |
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248 using collection as input (this is now the default input mode); |
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249 collect additional output dataset; disable ``usearch``-based |
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250 pipelines (i.e. ``UPARSE`` and ``QIIME``). |
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251 1.0.2 Enable support for FASTQs supplied via dataset collections and |
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252 fix some broken output datasets. |
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253 1.0.1 Initial version |
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254 ========== ====================================================================== |