annotate mqppep_anova.xml @ 31:c33c30398908 draft

planemo upload for repository https://github.com/galaxyproteomics/tools-galaxyp/tree/master/tools/mqppep commit 7e8b616a2519c8faeb6fd743a8fb73f271f655f9
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date Fri, 09 Dec 2022 21:05:54 +0000
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1 <tool
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2 id="mqppep_anova"
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3 name="MaxQuant Phosphopeptide ANOVA"
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4 version="@TOOL_VERSION@+galaxy@VERSION_SUFFIX@"
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5 profile="21.05"
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6 >
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7 <description>Runs ANOVA and KSEA for phosphopeptides.</description>
0
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8 <macros>
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9 <import>macros.xml</import>
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10 <xml name="group_matching_parm">
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11 <param name="group_filter_mode" type="select"
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12 help="Regular expression matching mode 'fixed', 'perl', or 'grep' with option for case insensitivity. See https://rdrr.io/r/base/grep.html"
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13 label="Sample-group matching mode"
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14 >
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15 <option value="r">ERE ("extended regular expressions")</option>
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16 <option value="ri"> - ERE, case insensitive</option>
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17 <option value="p" selected="true">PCRE ("PERL-compatible regular expressions")</option>
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18 <option value="pi"> - PCRE, case insensitive</option>
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19 <option value="f">fixed strings ("no regular expressions")</option>
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20 <option value="fi"> - fixed strings, case insensitive</option>
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21 </param>
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22 <param name="group_filter_patterns" type="text" value=".+"
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23 help="Comma-separated list of regular expressions matching group-names"
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24 label="Sample-group matching pattern">
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25 <sanitizer>
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26 <valid initial="string.printable">
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27 <remove value="&apos;"/>
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28 </valid>
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29 </sanitizer>
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30 </param>
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31 </xml>
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32 </macros>
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33 <edam_topics>
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34 <edam_topic>topic_0121</edam_topic><!-- proteomics -->
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35 <edam_topic>topic_3520</edam_topic><!-- proteomics experiment-->
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36 </edam_topics>
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37 <edam_operations>
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38 <edam_operation>operation_0276</edam_operation><!-- Analyse a network of protein interactions. -->
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39 <edam_operation>operation_0531</edam_operation><!-- Heat map generation -->
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40 <edam_operation>operation_2938</edam_operation><!-- Dendrogram generation -->
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41 <edam_operation>operation_2938</edam_operation><!-- Imputation -->
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42 <edam_operation>operation_3435</edam_operation><!-- Standardisation and normalisation -->
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43 <edam_operation>operation_3501</edam_operation><!-- Enrichment analysis -->
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44 <edam_operation>operation_3658</edam_operation><!-- Statistical inference -->
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45 </edam_operations>
7
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46 <expand macro="requirements"/>
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47 <!--
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48 The weird invocation used here is because knitr and install_tinytex
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49 both need access to a writeable directory, but most directories in a
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50 biocontainer are read-only, so this builds a pseudo-home under /tmp
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51 -->
31
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52 <required_files>
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53 <include path="KSEA_impl_flowchart.pdf" />
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54 <include path="kinase_name_uniprot_lut.tabular.bz2" />
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55 <include path="kinase_uniprot_description_lut.tabular.bz2" />
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56 <include path="kinase_uniprot_description_lut.tabular.bz2" />
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57 <include path="mqppep_anova.R" />
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58 <include path="mqppep_anova_preamble.tex" />
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59 <include path="mqppep_anova_script.Rmd" />
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60 <include path="perpage.tex" />
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61 </required_files>
0
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62 <command detect_errors="exit_code"><![CDATA[
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63 (printenv | sort) &&
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64 cp '$__tool_directory__/mqppep_anova_script.Rmd' . &&
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65 cp '$__tool_directory__/mqppep_anova.R' . &&
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66 cp '$__tool_directory__/kinase_name_uniprot_lut.tabular.bz2' . &&
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67 cp '$__tool_directory__/kinase_uniprot_description_lut.tabular.bz2' . &&
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68 cp '$__tool_directory__/mqppep_anova_preamble.tex' . &&
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69 cp '$__tool_directory__/perpage.tex' . &&
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70 cp '$__tool_directory__/KSEA_impl_flowchart.pdf' . &&
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71 Rscript mqppep_anova.R
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72 --inputFile '$input_file'
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73 --alphaFile '$alpha_file'
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74 --preproc_sqlite '$preproc_sqlite'
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75 --firstDataColumn '$intensity_column_regex_f'
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76 --imputationMethod $imputation.imputation_method
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77 #if $imputation.imputation_method == "random"
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78 --meanPercentile '$imputation.meanPercentile'
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79 --sdPercentile '$imputation.sdPercentile'
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80 #end if
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81 --regexSampleNames '$sample_names_regex_f'
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82 --regexSampleGrouping '$sample_grouping_regex_f'
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83 #if $group_filter.group_filter_method == "none"
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84 --sampleGroupFilter 'none'
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85 #else
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86 --sampleGroupFilter '$group_filter.group_filter_method'
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87 --sampleGroupFilterPatterns '$group_filter_patterns_f'
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88 --sampleGroupFilterMode '$group_filter.group_filter_mode'
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89 #end if
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90 --intensityMinValuesPerClass '$intnsty_min_vals_per_smpl_grp'
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91 --imputedDataFile '$imputed_data_file'
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92 --imputedQNLTDataFile '$imp_qn_lt_file'
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93 --ksea_sqlite '$ksea_sqlite'
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94 --kseaMinSubstrateCount '$ksea_min_substrate_count'
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95 --ksea_cutoff_threshold '$ksea_cutoff_threshold'
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96 --ksea_cutoff_statistic 'FDR'
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97 --kseaUseAbsoluteLog2FC '$ksea_use_absolute_log2_fc'
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98 --minQuality '$ksea_min_quality'
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99 --anova_ksea_metadata '$anova_ksea_metadata'
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100 --reportFile '$report_file'
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101 ]]></command>
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102 <!--
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103 -->
0
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104 <configfiles>
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105 <configfile name="sample_names_regex_f">
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106 $sample_names_regex
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107 </configfile>
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108 <configfile name="sample_grouping_regex_f">
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109 $sample_grouping_regex
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110 </configfile>
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111 <configfile name="group_filter_patterns_f">
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112 #if $group_filter.group_filter_method != "none"
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113 $group_filter.group_filter_patterns
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114 #end if
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115 </configfile>
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116 <configfile name="intensity_column_regex_f">
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117 $intensity_column_regex
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118 </configfile>
0
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119 </configfiles>
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120 <inputs>
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121 <!--
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122 needed inputs:
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123 - # should filters be used to identify sample-groups to be included or excluded
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124 sampleGroupFilter: !r c("none", "exclude", "include")[3]
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125 - # what patterns should be used to match sample-groups
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126 # (extracted by regexSampleGrouping) when determining sample-groups
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127 # that should be included or excluded
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128 sampleGroupFilterPatterns: ".*CR,N.*"
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129 - # minimum number of observed values per class
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130 intensityMinPerClass: 0
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131 - # what should be the primary criterion to eliminate excessive heatmap rows
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132 intensityHeatmapCriteria: !r c("quality", "na_count", "p_value")[1]
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133 suggested or advanced inputs:
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134 - kinaseNameUprtLutBz2: "./kinase_name_uniprot_lut.tabular.bz2"
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135 - kinaseUprtDescLutBz2: "./kinase_uniprot_description_lut.tabular.bz2"
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136 -->
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137 <param name="input_file" type="data" format="tabular" label="Filtered phosphopeptide intensities (tabular)"
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138 help="'preproc_tab' dataset produced by 'MaxQuant Phosphopeptide Preprocessing' tool"
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139 />
26
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140 <param name="alpha_file" type="data" format="tabular" label="ANOVA alpha cutoff level (tabular)"
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141 help="ANOVA alpha cutoff values for significance testing: tabular data having one column and no header"
0
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142 />
26
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143 <param name="preproc_sqlite" type="data" format="sqlite" label="Database from mqppep_preproc (sqlite)"
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144 help="'preproc_sqlite' dataset produced by 'MaxQuant Phosphopeptide Preprocessing' tool"
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145 />
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146 <param name="intensity_column_regex" type="text" value="^Intensity[^_]"
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147 label="Intensity-column pattern"
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148 help="Pattern matching columns that have peptide intensity data (PERL-compatible regular expression matching column label)"
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149 />
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150 <!-- imputation_method <- c("group-median","median","mean","random")[1] -->
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151 <conditional name="imputation">
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152 <param name="imputation_method" type="select" label="Imputation method"
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153 help="Impute missing values by (1) using median for each sample-group; (2) using median across all samples; (3) using mean across all samples; or (4) using randomly generated values having same SD as across all samples (with mean specified by 'Mean percentile for random values')"
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154 >
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155 <option value="random" selected="true">random</option>
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156 <option value="group-median">group-median</option>
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157 <option value="median">median</option>
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158 <option value="mean">mean</option>
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159 </param>
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160 <when value="group-median" />
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161 <when value="median" />
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162 <when value="mean" />
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163 <when value="random">
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164 <param name="meanPercentile" type="integer" value="1" min="1" max="99"
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165 label="Mean percentile for random values"
22
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166 help="Percentile center of random values; range [1,99]"
0
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167 />
26
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168 <param name="sdPercentile" type="float" value="1"
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169 label="Percentile SD for random values"
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170 help="Standard deviation adjustment-factor for random values; real number. (1.0 means SD of random values equal to the SD for the entire data set.)"
0
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171 />
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172 </when>
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173 </conditional>
12
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174 <param name="sample_names_regex" type="text" value="\.\d+[A-Z]$"
26
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175 help="Pattern extracting sample-names from names of columns of 'Filtered phosphopeptide intensities' that have peptide intensity data (PERL-compatible regular expression)"
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176 label="Sample-name extraction pattern">
0
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177 <sanitizer>
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178 <valid initial="string.printable">
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179 <remove value="&apos;"/>
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180 </valid>
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181 </sanitizer>
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182 </param>
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183 <param name="sample_grouping_regex" type="text" value="\d+"
26
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184 help="Pattern extracting sample-group from the extracted sample-names (PERL-compatible regular expression)"
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185 label="Sample-group extraction pattern">
0
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186 <sanitizer>
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187 <valid initial="string.printable">
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188 <remove value="&apos;"/>
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189 </valid>
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190 </sanitizer>
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191 </param>
26
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192 <param name="intnsty_min_vals_per_smpl_grp" type="integer" value="1" min="0"
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193 label="Minimum number of values per sample-group"
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194 help="Only consider as comparable those intensities having at least this number of values in each sample-group (range [0,&#8734;])"
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195 />
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196 <conditional name="group_filter">
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197 <param name="group_filter_method" type="select" label="Filter sample-groups"
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198 help="What filter should be applied to sample-group names? (1) 'none', no filter; (2) 'include', match is required; (3) 'exclude', match is forbidden."
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199 >
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200 <option value="none" selected="true">none</option>
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201 <option value="include">include</option>
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202 <option value="exclude">exclude</option>
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203 </param>
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204 <when value="none" />
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205 <when value="include">
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206 <expand macro="group_matching_parm"/>
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207 </when>
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208 <when value="exclude">
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209 <expand macro="group_matching_parm"/>
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210 </when>
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211 </conditional>
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212 <param name="ksea_min_substrate_count" type="integer" value="1" min="1"
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213 label="Minimum number of kinase-substrates for KSEA"
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214 help="Minimum number of substrates to consider any kinase for KSEA (range [1,&#8734;])"
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215 />
22
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216 <param name="ksea_cutoff_threshold" type="float" value="0.05"
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217 label="KSEA threshold level"
26
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218 help="Maximum FDR to be used to score a kinase enrichment as significant; see warning against setting this too low in help text below."
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219 />
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220 <param name="ksea_use_absolute_log2_fc"
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221 type="boolean"
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222 label="Use abs(log2(fold-change)) for KSEA"
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223 help="Should log2(fold-change) be used for KSEA? (Checking this may alter (possibly reduce) the number of hits.)"
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224 checked="false"
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225 truevalue="TRUE"
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226 falsevalue="FALSE"
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227 />
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228 <param name="ksea_min_quality" type="integer" value="0" min="0"
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229 label="Minimum quality of substrates for KSEA"
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230 help="Minimum 'quality' of substrates to be considered for KSEA (range [0,&#8734;]); higher numbers reduce the number of substrates considered - see help text below."
22
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231 />
0
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232 </inputs>
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233 <outputs>
26
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234 <!-- earlier outputs will appear lower in the history list; therefore, put report at the top -->
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235 <data name="ksea_sqlite" format="sqlite" label="${input_file.name}..${imputation.imputation_method}-imputed_ksea_sqlite" />
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236 <data name="anova_ksea_metadata" format="tabular" label="${input_file.name}.${imputation.imputation_method}-anova_ksea_metadata" />
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237 <data name="imputed_data_file" format="tabular" label="${input_file.name}.${imputation.imputation_method}-imputed_intensities" />
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238 <data name="imp_qn_lt_file" format="tabular" label="${input_file.name}.${imputation.imputation_method}-imputed_QN_LT_intensities" />
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239 <data name="report_file" format="pdf" label="${input_file.name}.${imputation.imputation_method}-imputed_report" />
0
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240 </outputs>
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241 <tests>
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242 <test><!-- test #1 -->
0
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243 <param name="input_file" ftype="tabular" value="test_input_for_anova.tabular"/>
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244 <param name="preproc_sqlite" ftype="sqlite" value="test_input_for_anova.sqlite"/>
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245 <param name="alpha_file" ftype="tabular" value="alpha_levels.tabular"/>
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246 <param name="intensity_column_regex" value="^Intensity[^_]"/>
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247 <param name="imputation_method" value="median"/>
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248 <param name="sample_names_regex" value="\.\d+[A-Z]$"/>
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249 <param name="sample_grouping_regex" value="\d+"/>
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250 <output name="imputed_data_file">
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251 <assert_contents>
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252 <has_text text="Phosphopeptide" />
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253 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
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254 <!-- missing missing observd missing observd observd -->
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255 <has_text_matching expression="pSQKQEEENPAEETGEEK.*8765300.8765300.8765300.8765300.2355900.14706000" />
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256
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257 </assert_contents>
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258 </output>
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259 <output name="imp_qn_lt_file">
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260 <assert_contents>
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261 <has_text text="Phosphopeptide" />
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262 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
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263 <!-- missing missing observed missing observed observed -->
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264 <has_text_matching expression="pSQKQEEENPAEETGEEK.*6.962256.*6.908828.*6.814580.*6.865411.*6.908828.*7.093748" />
22
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265
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266 <has_text text="pSQKQEEENPAEETGEEK" />
0
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267 </assert_contents>
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268 </output>
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269 </test>
26
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270 <test><!-- test #2 -->
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271 <param name="input_file" ftype="tabular" value="test_input_for_anova.tabular"/>
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272 <param name="preproc_sqlite" ftype="sqlite" value="test_input_for_anova.sqlite"/>
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273 <param name="alpha_file" ftype="tabular" value="alpha_levels.tabular"/>
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274 <param name="intensity_column_regex" value="^Intensity[^_]"/>
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275 <param name="imputation_method" value="mean"/>
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276 <!--
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277 <param name="meanPercentile" value="1"/>
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278 <param name="sdPercentile" value="1"/>
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279 -->
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280 <param name="sample_names_regex" value="\.\d+[A-Z]$"/>
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281 <param name="sample_grouping_regex" value="\d+"/>
26
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282 <param name="intnsty_min_vals_per_smpl_grp" value="1"/>
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283 <param name="group_filter_method" value="none"/>
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284 <!--
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285 <param name="group_filter_mode" value="r"/>
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286 <param name="group_filter_patterns" value="\.+"/>
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287 -->
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288 <param name="ksea_min_substrate_count" value="1"/>
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289 <param name="ksea_cutoff_threshold" value="0.5"/>
22
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290 <output name="imputed_data_file">
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291 <assert_contents>
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292 <has_text text="Phosphopeptide" />
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293 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
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294 <!-- missing missing observd missing observd observd -->
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295 <has_text_matching expression="pSQKQEEENPAEETGEEK.*6721601.6721601.8765300.6721601.2355900.14706000" />
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296
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297 </assert_contents>
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298 </output>
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299 <output name="imp_qn_lt_file">
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300 <assert_contents>
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301 <has_text text="Phosphopeptide" />
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302 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
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303 <!-- missing missing observed missing observed observed -->
26
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304 <has_text_matching expression="pSQKQEEENPAEETGEEK.*6.839850.*6.797424.*6.797424.*6.797424.*6.896609.*7.097251" />
22
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305 </assert_contents>
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306 </output>
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307 </test>
26
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308 <test><!-- test #3 -->
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309 <param name="input_file" ftype="tabular" value="test_input_for_anova.tabular"/>
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310 <param name="preproc_sqlite" ftype="sqlite" value="test_input_for_anova.sqlite"/>
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311 <param name="alpha_file" ftype="tabular" value="alpha_levels.tabular"/>
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312 <param name="intensity_column_regex" value="^Intensity[^_]"/>
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313 <param name="imputation_method" value="group-median"/>
26
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314 <!--
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315 <param name="meanPercentile" value="1"/>
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316 <param name="sdPercentile" value="1"/>
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317 -->
22
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318 <param name="sample_names_regex" value="\.\d+[A-Z]$"/>
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319 <param name="sample_grouping_regex" value="\d+"/>
26
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320 <param name="intnsty_min_vals_per_smpl_grp" value="1"/>
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321 <param name="group_filter_method" value="none"/>
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322 <!--
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323 <param name="group_filter_mode" value="r"/>
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324 <param name="group_filter_patterns" value="\.+"/>
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325 -->
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326 <param name="ksea_min_substrate_count" value="1"/>
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327 <param name="ksea_cutoff_threshold" value="0.5"/>
22
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328 <output name="imputed_data_file">
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329 <assert_contents>
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330 <has_text text="Phosphopeptide" />
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331 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
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332 <!-- missing missing observd missing observd observd -->
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333 <has_text_matching expression="pSQKQEEENPAEETGEEK.*8765300.8765300.8765300.5886074.2355900.14706000" />
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334
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335 </assert_contents>
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336 </output>
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337 <output name="imp_qn_lt_file">
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338 <assert_contents>
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339 <has_text text="Phosphopeptide" />
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340 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
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341 <!-- missing missing observed missing observed observed -->
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342 <has_text_matching expression="pSQKQEEENPAEETGEEK.*6.946112.*6.888985.*6.792137.*6.792137.*6.888985.*7.089555" />
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343 </assert_contents>
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344 </output>
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345 </test>
26
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346 <test><!-- test #4 -->
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347 <param name="input_file" ftype="tabular" value="test_input_for_anova.tabular"/>
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348 <param name="preproc_sqlite" ftype="sqlite" value="test_input_for_anova.sqlite"/>
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349 <param name="alpha_file" ftype="tabular" value="alpha_levels.tabular"/>
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350 <param name="intensity_column_regex" value="^Intensity[^_]"/>
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351 <param name="imputation_method" value="random"/>
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352 <param name="meanPercentile" value="1" />
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353 <param name="sdPercentile" value="1.0" />
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354 <param name="sample_names_regex" value="\.\d+[A-Z]$"/>
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355 <param name="sample_grouping_regex" value="\d+"/>
22
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356 <output name="imputed_data_file">
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357 <assert_contents>
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358 <has_text text="Phosphopeptide" />
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359 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
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360 <!-- observd observd observd -->
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361 <has_text_matching expression="pSQKQEEENPAEETGEEK.*8765300.*2355900.*4706000" />
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362
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363 </assert_contents>
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364 </output>
13
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365 <output name="imp_qn_lt_file">
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366 <assert_contents>
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367 <has_text text="Phosphopeptide" />
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368 <has_text text="AAAITDMADLEELSRLpSPLPPGpSPGSAAR" />
26
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369 <has_text text="5.522821" /> <!-- log-transformed value for pTYVDPFTpYEDPNQAVR .1B -->
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370 <has_text text="6.638251" /> <!-- log-transformed value for pSQKQEEENPAEETGEEK .2A -->
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371 </assert_contents>
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372 </output>
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373 </test>
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374 </tests>
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375 <help><![CDATA[
22
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376 ====================================================
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377 Phopsphoproteomic Enrichment Pipeline ANOVA and KSEA
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378 ====================================================
0
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379
26
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380 **Overview**
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381 ============
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382
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383 Perform statistical analysis of preprocessed MaxQuant output data collected as described in `[Cheng, 2018] <https://doi.org/10.3791/57996>`_.
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384
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385 - Extracts sample-group IDs from sample names.
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386 - Imputes missing values.
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387 - Performs ANOVA analysis for each phosphopeptide.
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388 - Performs Kinase-Substrate Enrichment Analysis (KSEA) using the method described by `Casado et al. (2013) <doi:10.1126/scisignal.2003573>`_; see *"Algorithms"* section below.
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389
26
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390 **Workflow position**
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391 =====================
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392
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393 Upstream tool
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394 The "MaxQuant Phosphopeptide Preprocessing" tool (``mqppep_preproc``) that transforms MaxQuant output for phospoproteome-enriched samples into a form suitable for statistical analysis.
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395
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396 **Input datasets**
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397 ==================
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398
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399 ``Filtered phosphopeptide intensities`` (tabular)
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400 Phosphopeptides annotated with SwissProt and phosphosite metadata (in tabular format).
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401 This is the output from the "MaxQuant Phopsphopeptide Preprocessing"
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402 (``mqppep_preproc``) tool.
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403
26
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404 - First column label 'Phosphopeptide'.
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405 - Sample-intensities must begin in first column matching 'Intensity-column pattern' and must have column labels to match argument 'Sample-name extraction pattern'.
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406
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407 ``ANOVA alpha cutoff level`` (tabular)
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408 List of alpha cutoff values for significance testing; text file having one column and no header. For example:
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409
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410 ::
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411
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412 0.2
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413 0.1
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414 0.05
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415
26
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416 ``Database from mqppep_preproc`` (sqlite)
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417 SQLite database produced by the "MaxQuant Phopsphopeptide Preprocessing"
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418 (``mqppep_preproc``) tool.
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419
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420 **Input parameters**
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421 ====================
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422
23
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423 ``Intensity-column pattern``
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424 First column of ``Filtered phosphopeptide intensities`` having intensity values (integer or PERL-compatible regular expression matching column label). Default::
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425
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426 ^Intensity[^_]
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427
23
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428 ``Imputation method``
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429 Impute missing values by:
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430
26
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431 1. ``group-median`` - use median for each sample-group;
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432 2. ``mean`` - use mean across all samples; or
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433 3. ``median`` - use median across all samples;
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434 4. ``random`` - use randomly generated values where:
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435
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436 (i) ``Mean percentile for random values`` specifies the percentile among non-missing values to be used as mean of random values, and
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437 (ii) ``Percentile SD for random values`` specifies the factor to be multiplied by the standard deviation among the non-missing values (across all samples) to determine the standard deviation of random values.
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438
26
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439 ``Sample-name extraction pattern``
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440 PERL-compatible regular expression extracting the sample-name from the the name of a column of intensities (from ``Filtered phosphopeptide intensities``) for one sample.
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441
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442 - For example, ``"\.\d+[A-Z]$"`` applied to "``Intensity.splunge.10A``" would produce "``.10A``".
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443 - Note that *this is case sensitive* by default.
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444
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445 ``Sample-group extraction pattern``
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446 PERL-compatible regular expression extracting the sample-grouping from the sample-name (that was in turn extracted with ``Sample-name extraction pattern`` from a column of intensites from ``Filtered phosphopeptide intensities``).
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447
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448 - For example, ``"\d+$"`` applied to "``.10A``" would produce "``10``".
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449 - Note that *this is case sensitive* by default.
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450
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451 ``Minimum number of values per sample-group``
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452 Sometimes you may wish to filter out the intensities that are poorly represented among some sample groups because they complicate the comparison process. You can use this parameter to specify the minimum number of values in any sample-group (range [0,]]>&#8734;<![CDATA[])
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453
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454 ``Filter sample-groups``
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455 Sometimes you may have spectra that are for treatments that you are not considering for your comparison. You can specify a filter (or not) for sample-group names; if you do, you can specify whether groups that match your criteria should be excluded from the analysis ("forbidden") or included in the analysis ("required").
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456
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457 ``Sample-group matching mode``
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458 The R `base::grep` function that is used here for pattern matching is exhaustively documented at https://rdrr.io/r/base/grep.html. There are two choices you make here. The first is whether to differentiate lowercase and uppercase characters. The second is wheter to require exact matches ("fixed" pattern-matching mode) or to use "PERL-compatible regular expressions) ("perl") or "extendd regular expressions" ("grep"). See https://rdrr.io/r/base/grep.html for further info.
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459
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460 ``Sample-group matching pattern``
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461 This is a comma-separated list of patterns to match to group-names, according to the ``Sample-group matching mode`` that you have chosen.
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462
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463 ``Minimum number of kinase-substrates for KSEA``
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464 For KSEA, you may decide that you wish to ignore kinases having fewer substrates than some minimum; specify that minimum here (range [1,]]>&#8734;<![CDATA[])
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465
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466 ``KSEA threshold level``
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467 Specifies minimum FDR at which a kinase will be considered to be enriched; the default choice of ``0.05`` is arbitrary and may exclude kinases that are interesting. The KSEA FDR perhaps should not be treated as conservatively as would be appropriate for hypothesis testing. For example, at an FDR of ``0.05``, for every ``20`` kinases that on discards, ``19`` are likely truely enriched.
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468
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469 ``Use abs(log2(fold-change)) for KSEA``
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470 When TRUE, consider only the magnitude of the differences across the contrast for all of the substrates when aggregating them to assess the enrichment of a given kinase's substrates. When FALSE, also consider the direction. Surprisingly, setting this to TRUE may decrease the enriched kinases.
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471
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472 ``Minimum quality of substrates for KSEA``
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473 An arbitrary "quality score" is assigned to each substrate, as described in the PDF report produced by the tool. This score takes into account both FDR-adjusted p-value and the number of missing values for each substrate. Setting the minimum to zero retains all substrates, which may be a large number.
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474
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475 **Outputs**
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476 ===========
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477
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478 Report dataset
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479 *[input file].[imputation method]*-``imputed_report``
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480
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481 Summary report for normalization, imputation, and **ANOVA**, in PDF format.
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482
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483 Imputed intensities
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484 *[input file].[imputation method]*-``imputed_intensities``
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485
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486 Phosphopeptide MS intensities where missing values have been **imputed** by the chosen method, in tabular format.
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487
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488 Imputed quantum-normalized log-transformed intensities
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489 *[input file].[imputation method]*-``imputed_QN_LT_intensities``
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490
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491 Phosphopeptide MS intensities where missing values have been **imputed** by the chosen method, quantile-normalized (**QN**), and log10-transformed (**LT**), in tabular format.
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492
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493 ANOVA KSEA metadata
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494 *[input file].[imputation method]*-``imputed_anova_ksea_metadata``
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495 Phosphopeptide metadata including ANOVA significance and KSEA enrichments.
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496
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497 KSEA SQLite database sqlite
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498 *[input file].[imputation method]*-``imputed_ksea_sqlite``
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499 An SQLite database that is usable for *ad hoc* report creation.
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500
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501 **Algorithm**
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502 =============
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503
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504 The KSEA algorithm used here is as in the KSEAapp package as reported in `[Wiredja 2017] <https://doi.org/10.1093/bioinformatics/btx415>`_.
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505 The code is adapted from `"Danica D. Wiredja (2017). KSEAapp: Kinase-Substrate Enrichment Analysis. R package version 0.99.0." <https://cran.r-project.org/package=KSEAapp>`_ to work with output from the "MaxQuant Phosphopeptide Preprocessing" Galaxy tool and the multiple kinase-substrate databases that the latter tool searches.
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506
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507 **Authors**
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508 ===========
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509
c1403d18c189 "planemo upload for repository https://github.com/galaxyproteomics/tools-galaxyp/tree/master/tools/mqppep commit bb6c941be50db4c0719efdeaa904d7cb7aa1d182"
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510 ``Larry C. Cheng``
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511 (`ORCiD 0000-0002-6922-6433 <https://orcid.org/0000-0002-6922-6433>`_) wrote the original script.
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512
c1403d18c189 "planemo upload for repository https://github.com/galaxyproteomics/tools-galaxyp/tree/master/tools/mqppep commit bb6c941be50db4c0719efdeaa904d7cb7aa1d182"
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513 ``Arthur C. Eschenlauer``
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514 (`ORCiD 0000-0002-2882-0508 <https://orcid.org/0000-0002-2882-0508>`_) adapted the script to run in Galaxy.
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515
c1403d18c189 "planemo upload for repository https://github.com/galaxyproteomics/tools-galaxyp/tree/master/tools/mqppep commit bb6c941be50db4c0719efdeaa904d7cb7aa1d182"
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516 ===================================
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517 PERL-compatible regular expressions
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518 ===================================
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519
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520 Note that the PERL-compatible regular expressions accepted by this tool are documented at http://rdrr.io/r/base/regex.html
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521
c1403d18c189 "planemo upload for repository https://github.com/galaxyproteomics/tools-galaxyp/tree/master/tools/mqppep commit bb6c941be50db4c0719efdeaa904d7cb7aa1d182"
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522 ]]></help>
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523 <citations>
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524 <!-- Cheng_2018 "Phosphopeptide Enrichment ..." PMID: 30124664 -->
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525 <citation type="doi">10.3791/57996</citation>
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526 <!-- Wiredja_2017 "The KSEA App ..." PMID: 28655153 -->
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527 <citation type="doi">10.1093/bioinformatics/btx415</citation>
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528 <citation type="bibtex">@Manual{,
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529 title = {KSEAapp: Kinase-Substrate Enrichment Analysis},
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530 author = {Danica D. Wiredja},
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531 year = {2017},
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532 note = {R package version 0.99.0},
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533 }</citation>
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534 </citations>
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535 </tool>