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1 <tool id="bismark_methylation_extractor" name="Bismark Meth. Extractor" version="0.10.1">
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2 <!-- Wrapper compatible with Bismark version 0.10 -->
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3 <description>Reports on methylation status of reads mapped by Bismark</description>
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0
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4 <!--<version_command>bismark_methylation_extractor version</version_command>-->
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5 <requirements>
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6 <requirement type="set_environment">SCRIPT_PATH</requirement>
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7 <requirement type="package" version="0.12.8">bowtie</requirement>
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3
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8 <requirement type="package" version="2.1.0">bowtie2</requirement>
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9 </requirements>
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10 <parallelism method="basic"></parallelism>
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11 <command interpreter="python">
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12 bismark_methylation_extractor.py
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13
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14 --infile $input
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15
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16 --bismark_path \$SCRIPT_PATH
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17
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18 #if $singlePaired.sPaired == "single":
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19 --single-end
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20 #else:
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21 --paired-end
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22 $singlePaired.no_overlap
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23 #end if
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24
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25 #if str($ignore_bps) != "0":
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26 --ignore $ignore_bps
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27 #end if
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28
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29 #if $report:
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30 --report-file $o_report
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31 #end if
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32
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33 #if $comprehensive:
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34 --comprehensive
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35 #end if
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36
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37 #if $merge_non_cpg:
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38 --merge-non-cpg
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39 #end if
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40
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41 #if $compress:
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42 --compress $compressed_output
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43 #else:
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44 #if $comprehensive == False and $merge_non_cpg == False:
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45 ##twelfe files
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46 --cpg_ot $cpg_ot
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47 --chg_ot $chg_ot
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48 --chh_ot $chh_ot
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49 --cpg_ctot $cpg_ctot
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50 --chg_ctot $chg_ctot
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51 --chh_ctot $chh_ctot
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52 --cpg_ob $cpg_ob
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53 --chg_ob $chg_ob
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54 --chh_ob $chh_ob
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55 --cpg_ctob $cpg_ctob
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56 --chg_ctob $chg_ctob
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57 --chh_ctob $chh_ctob
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58 #elif $merge_non_cpg and $comprehensive:
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59 ## two files
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60 --non_cpg_context $non_cpg_context
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61 --cpg_context $cpg_context
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62 #elif $comprehensive:
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63 ## three files
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64 --cpg_context $cpg_context
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65 --chg_context $chg_context
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66 --chh_context $chh_context
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67 #elif $merge_non_cpg:
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68 ## eight files
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69 --non_cpg_context_ctot $non_cpg_context_ctot
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70 --non_cpg_context_ot $non_cpg_context_ot
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71 --non_cpg_context_ob $non_cpg_context_ob
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72 --non_cpg_context_ctob $non_cpg_context_ctob
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73 --cpg_ot $cpg_ot
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74 --cpg_ctot $cpg_ctot
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75 --cpg_ob $cpg_ob
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76 --cpg_ctob $cpg_ctob
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77 #end if
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78 ## end compress
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79 #end if
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80
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81 </command>
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82 <inputs>
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83 <!-- Input Parameters -->
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84 <param name="input" type="data" format="sam,bam" label="SAM/BAM file from Bismark bisulfite mapper" />
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85 <conditional name="singlePaired">
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86 <param name="sPaired" type="select" label="Is this library mate-paired?">
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87 <option value="single">Single-end</option>
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88 <option value="paired">Paired-end</option>
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89 </param>
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90 <when value="single" />
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91 <when value="paired">
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92 <param name="no_overlap" type="boolean" truevalue="--no-overlap" falsevalue="" checked="False" label="This option avoids scoring overlapping methylation calls twice, in case of overlapping read one and read two" help="" />
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93 </when>
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94 </conditional>
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95 <param name="ignore_bps" type="integer" value="0" label="Ignore the first N bp when processing the methylation call string" />
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96 <param name="comprehensive" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Merge all four possible strand-specific methylation info
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97 into context-dependent output files" help="" />
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98 <param name="merge_non_cpg" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Merge all non-CpG contexts into one file" help="This will produce eight strand-specific output files, or two output files in comprehensive mode." />
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99 <param name="report" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Short methylation summary output" />
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100 <param name="compress" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Compress all result files and output one single file" />
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101
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102 </inputs>
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103 <outputs>
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104 <!--
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105 OT – original top strand
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106 CTOT – complementary to original top strand
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107 OB – original bottom strand
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108 CTOB – complementary to original bottom strand
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109 -->
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110 <data format="tabular" name="o_report" label="${tool.name} on ${on_string}: Report file">
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111 <filter> ( report is True ) </filter>
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112 </data>
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113
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114 <!-- default output 12 files -->
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115 <data format="tabular" name="cpg_ot" label="${tool.name} on ${on_string}: CpG original top strand">
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116 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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117 </data>
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118 <data format="tabular" name="chg_ot" label="${tool.name} on ${on_string}: CHG original top strand">
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119 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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120 </data>
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121 <data format="tabular" name="chh_ot" label="${tool.name} on ${on_string}: CHH original top strand">
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122 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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123 </data>
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124 <data format="tabular" name="cpg_ctot" label="${tool.name} on ${on_string}: CpG complementary to top strand">
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125 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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126 </data>
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127 <data format="tabular" name="chg_ctot" label="${tool.name} on ${on_string}: CHG complementary to top strand">
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128 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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129 </data>
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130 <data format="tabular" name="chh_ctot" label="${tool.name} on ${on_string}: CHH complementary to top strand">
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131 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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132 </data>
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133
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134 <data format="tabular" name="cpg_ob" label="${tool.name} on ${on_string}: CpG original bottom strand">
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135 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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136 </data>
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137 <data format="tabular" name="chg_ob" label="${tool.name} on ${on_string}: CHG original bottom strand">
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138 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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139 </data>
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140 <data format="tabular" name="chh_ob" label="${tool.name} on ${on_string}: CHH original bottom strand">
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141 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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142 </data>
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143 <data format="tabular" name="cpg_ctob" label="${tool.name} on ${on_string}: CpG complementary to bottom strand">
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144 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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145 </data>
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146 <data format="tabular" name="chg_ctob" label="${tool.name} on ${on_string}: CHG complementary to bottom strand">
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147 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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148 </data>
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149 <data format="tabular" name="chh_ctob" label="${tool.name} on ${on_string}: CHH complementary to bottom strand">
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150 <filter> ( compress == False and comprehensive == False and merge_non_CpG == False) </filter>
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151 </data>
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152
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153 <!-- Context-dependent methylation output files (comprehensive option) -->
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154 <data format="tabular" name="cpg_context" label="${tool.name} on ${on_string}: CpG context dependent">
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155 <filter> ( compress == False and comprehensive) </filter>
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156 </data>
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157 <data format="tabular" name="chg_context" label="${tool.name} on ${on_string}: CHG context dependent">
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158 <filter> ( compress == False and comprehensive and merge_non_CpG == False) </filter>
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159 </data>
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160 <data format="tabular" name="chh_context" label="${tool.name} on ${on_string}: CHH context dependent">
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161 <filter> ( compress == False and comprehensive and merge_non_CpG == False) </filter>
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162 </data>
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163
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164 <data format="tabular" name="non_cpg_context" label="${tool.name} on ${on_string}: Non CpG context dependent">
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165 <filter> ( compress == False and comprehensive and merge_non_cpg) </filter>
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166 </data>
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167
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168 <data format="tabular" name="non_cpg_context_ot" label="${tool.name} on ${on_string}: Non CpG context dependent on original top strand">
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169 <filter> ( compress == False and comprehensive == False and merge_non_cpg) </filter>
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170 </data>
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171 <data format="tabular" name="non_cpg_context_ctot" label="${tool.name} on ${on_string}: Non CpG context dependent on complementary to top strand">
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172 <filter> ( compress == False and comprehensive == False and merge_non_cpg) </filter>
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173 </data>
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174 <data format="tabular" name="non_cpg_context_ob" label="${tool.name} on ${on_string}: Non CpG context dependent on bottom top strand">
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175 <filter> ( compress == False and comprehensive == False and merge_non_cpg) </filter>
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176 </data>
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177 <data format="tabular" name="non_cpg_context_ctob" label="${tool.name} on ${on_string}: Non CpG context dependent on complementary to bottom strand">
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178 <filter> ( compress == False and comprehensive == False and merge_non_cpg) </filter>
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179 </data>
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180
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181 <data format="gzipped" name="compressed_output" label="${tool.name} on ${on_string}: Result archive.">
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182 <filter> ( compress ) </filter>
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183 </data>
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184 </outputs>
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185
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186 <tests>
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187 </tests>
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188
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189 <help>
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190
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191 **What it does**
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192
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193 The following is a brief description of all options to control the Bismark_
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194 methylation extractor. The script reads in a bisulfite read alignment results file
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195 produced by the Bismark bisulfite mapper and extracts the methylation information
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196 for individual cytosines. This information is found in the methylation call field
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197 which can contain the following characters:
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198
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199
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200 - X = for methylated C in CHG context (was protected)
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201 - x = for not methylated C CHG (was converted)
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202 - H = for methylated C in CHH context (was protected)
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203 - h = for not methylated C in CHH context (was converted)
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204 - Z = for methylated C in CpG context (was protected)
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205 - z = for not methylated C in CpG context (was converted)
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206 - . = for any bases not involving cytosines
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207
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208
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209 The methylation extractor outputs result files for cytosines in CpG, CHG and CHH
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210 context (this distinction is actually already made in Bismark itself). As the methylation
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211 information for every C analysed can produce files which easily have tens or even hundreds of
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212 millions of lines, file sizes can become very large and more difficult to handle. The C
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213 methylation info additionally splits cytosine methylation calls up into one of the four possible
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214 strands a given bisulfite read aligned against:
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215
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216 - OT = original top strand
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217 - CTOT = complementary to original top strand
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218
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219 - OB = original bottom strand
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220 - CTOB = complementary to original bottom strand
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221
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222 Thus, by default twelve individual output files are being generated per input file (unless
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223 --comprehensive is specified, see below). The output files can be imported into a genome
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224 viewer, such as SeqMonk, and re-combined into a single data group if desired (in fact
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225 unless the bisulfite reads were generated preserving directionality it doesn't make any
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226 sense to look at the data in a strand-specific manner). Strand-specific output files can
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227 optionally be skipped, in which case only three output files for CpG, CHG or CHH context
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228 will be generated. For both the strand-specific and comprehensive outputs there is also
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229 the option to merge both non-CpG contexts (CHG and CHH) into one single non-CpG context.
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230
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231
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232 .. _Bismark: http://www.bioinformatics.babraham.ac.uk/projects/bismark/
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233
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234
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235 It is developed by Krueger F and Andrews SR. at the Babraham Institute. Krueger F, Andrews SR. (2011) Bismark: a flexible aligner and methylation caller for Bisulfite-Seq applications. Bioinformatics, 27, 1571-2.
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236
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237 -------
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238
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239 **Bismark settings**
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240
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241 All of the options have a default value. You can change any of them. If any Bismark function is missing please contact the tool author or your Galaxy admin.
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242
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243 ------
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244
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245 **Outputs**
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246
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247 The output files are in the following format (tab delimited)::
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248
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249
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250 Column Description
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251 -------- --------------------------------------------------------
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252 1 seq-ID
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253 2 strand
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254 3 chromosome
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255 4 position
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256 5 methylation call
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257
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258
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259 * Methylated cytosines receive a '+' orientation,
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260 * Unmethylated cytosines receive a '-' orientation.
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261
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262 ------
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263
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264 **OPTIONS**
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265
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266 Input::
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267
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268 -s/--single-end Input file(s) are Bismark result file(s) generated from single-end
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269 read data. Specifying either --single-end or --paired-end is
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270 mandatory.
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271
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272 -p/--paired-end Input file(s) are Bismark result file(s) generated from paired-end
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273 read data. Specifying either --paired-end or --single-end is
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274 mandatory.
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275
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276 --no_overlap For paired-end reads it is theoretically possible that read_1 and
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277 read_2 overlap. This option avoids scoring overlapping methylation
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278 calls twice. Whilst this removes a bias towards more methylation calls
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279 towards the center of sequenced fragments it can de facto remove
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280 a good proportion of the data.
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281
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282 --ignore INT Ignore the first INT bp at the 5' end of each read when processing the
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283 methylation call string. This can remove e.g. a restriction enzyme site
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284 at the start of each read.
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285
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286 Output::
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287
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288 --comprehensive Specifying this option will merge all four possible strand-specific
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289 methylation info into context-dependent output files. The default
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290 contexts are:
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291 - CpG context
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292 - CHG context
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293 - CHH context
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294
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295 --merge_non_CpG This will produce two output files (in --comprehensive mode) or eight
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296 strand-specific output files (default) for Cs in
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297 - CpG context
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298 - non-CpG context
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299
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300 --report Prints out a short methylation summary as well as the paramaters used to run
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301 this script.
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302
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303
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304 </help>
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305 </tool>
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