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12
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1 <tool id="tarean" name="Tandem Repeat Analyzer" version="2.3.8.1" >
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2
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2 <stdio>
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3 <regex match="Traceback" source="stderr" level="fatal" description="Unknown error" />
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4 <regex match="error" source="stderr" level="fatal" description="Unknown error" />
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5 <regex match="warning" source="stderr" level="warning" description="Unknown warning" />
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6 <exit_code range="1:" level="fatal" description="Error" />
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7 </stdio>
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8 <description>Identification of genomic tandem repeats from NGS data</description>
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4
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9 <requirements>
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6
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10 <container type="singularity">library://repeatexplorer/default/repex_tarean:0.3.8-dbaa07f</container>
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4
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11 </requirements>
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2
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12 <command detect_errors="exit_code">
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13 export PYTHONHASHSEED=0;
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14 seqclust --paired --sample ${read_sampling.sample} --output_dir=tarean_output --logfile=${log} --cleanup --tarean_mode
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15 #if $advanced_options.advanced:
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16 --mincl $advanced_options.size_threshold $advanced_options.keep_names $advanced_options.automatic_filtering -M $advanced_options.merging
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17 #if $advanced_options.custom_library.options_custom_library :
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18 -d $advanced_options.custom_library.library extra_database
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19 #end if
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20 #if $advanced_options.options.options:
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21 -opt $advanced_options.options.options
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22 #end if
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23 #else:
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24 -M 0.2
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25
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26 #end if
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27 ${FastaFile} >stdout.log 2> stderr.log ;
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28 echo "STDOUT CONTENT:" >> ${log} ;
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29 cat stdout.log >> ${log} ;
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30 echo "STDERR CONTENT:" >> ${log} ;
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31 cat stderr.log >> ${log} &&
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32 /opt/repex_tarean/stderr_filter.py stderr.log &&
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2
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33 cd tarean_output &&
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34 zip -r ${ReportArchive}.zip * &&
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35 mv ${ReportArchive}.zip ${ReportArchive} &&
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36 cp index.html ${ReportFile} &&
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37 mkdir -p ${ReportFile.extra_files_path} &&
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38 cp -r --parents libdir ${ReportFile.extra_files_path} &&
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39 cp -r --parents seqclust/clustering/superclusters ${ReportFile.extra_files_path} &&
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40 cp -r --parents seqclust/clustering/clusters ${ReportFile.extra_files_path} &&
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41 cp seqclust/clustering/hitsort.cls ${ReportFile.extra_files_path}/seqclust/clustering/hitsort.cls &&
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42 cp *.png ${ReportFile.extra_files_path}/ &&
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43 cp *.csv ${ReportFile.extra_files_path}/ &&
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44 cp *.html ${ReportFile.extra_files_path}/ &&
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45 cp *.css ${ReportFile.extra_files_path}/ &&
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46 cp *.fasta ${ReportFile.extra_files_path}/ 2>>$log && rm -r ../tarean_output || :
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2
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47
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48
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49 </command>
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50
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51 <inputs>
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52 <param name="FastaFile" label="Paired-end Illumina reads" type="data" format="fasta"
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53 help="Input file must contain FASTA-formatted interlaced read pairs from paired-end sequencing. All pairs must be complete. Example of the input data format is provided in the help below."/>
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54
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55 <conditional name="read_sampling">
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56 <param name="do_sampling" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Read sampling" help="Use this option if you want to analyze only a part of the reads" />
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57 <when value="false">
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58 <!-- pass -->
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59 <param name="sample" label="Sample size" hidden="True" type="integer" value="0" help="Number of analyzed reads"/>
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60 </when>
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61 <when value="true">
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62 <param name="sample" label="Sample size" type="integer" value="500000" min="10000" help="Number of analyzed reads"/>
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63 </when>
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64 </conditional>
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65
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66 <conditional name="advanced_options">
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67 <param name="advanced" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Advanced options" />
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68 <when value="false">
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69 <!-- pass -->
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70 </when>
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71 <when value="true">
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72 <param name="merging" type="boolean" truevalue="0.2" falsevalue="0" checked="True" label="Perform cluster merging" help="By default, clusters connected through paired-end reads are merged"/>
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73 <conditional name="custom_library">
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74 <param name="options_custom_library" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Use custom repeat database"/>
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75 <when value="false">
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76 <!-- do nothing here -->
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77 </when>
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78 <when value="true">
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79 <param name="library" format="fasta" type="data" label="Use custom repeat database" help="Perform additional similarity search to user-provided repeat database. The database should contain FASTA-formatted DNA sequences with headers (sequence names) in the format: '>reapeatname#class/subclass'"/>
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80 </when>
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81 </conditional>
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82 <param name="size_threshold" label="Cluster size threshold for detailed analysis" type="float" value="0.01" min="0.0001" max="100" help ="Minimal size (as percentage of input reads) of the smallest cluster which is analyzed; cluster with less than 20 reads are not considered."/>
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83 <param name="automatic_filtering" label="Perform automatic filtering of abundant satellite repeats" type="boolean" truevalue="--automatic_filtering" falsevalue="" checked="false"/>
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84 <param name="keep_names" label="Keep original read names" type="boolean" truevalue="--keep_names" falsevalue="" checked="false" help="By default, reads are renamed using integers. Use this option if you want to keep original names."/>
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85 <conditional name="options">
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86 <param name="options" type="select" label="Similarity search options">
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87 <option value="ILLUMINA" selected="true">Default </option>
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88 <option value="ILLUMINA_DUST_OFF" selected="false">Masking of low complexity repeats disabled </option>
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89
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90 <!-- <option value="ILLUMINA_SENSITIVE_MGBLAST" selected="false">Illumina reads, sensitive search (search parameters: mgblast, min PID 80, -W8) slow, experimental feature!</option> -->
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91 <!-- <option value="ILLUMINA_SENSITIVE_BLASTPLUS" selected="false">Illumina reads, more sensitive search (search parameters: blastn, min PID 80, -W6) extremely slow, experimental feature!</option> -->
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92 <!-- <option value="OXFORD_NANOPORE" selected="false"> -->
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93 <!-- Pseudo short reads simulated from Oxford Nanopore data, experimental feature! -->
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94 <!-- </option> -->
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95 </param>
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96 </conditional>
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97 </when>
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98 </conditional>
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99
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100 <conditional name="queue_definition">
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101 <param name="queue_select" type="select" label="Select queue">
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102 <option value="basic_fast_queue">basic (max runtime 2 days, 4 GB RAM)</option>
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103 <option value="long_slow_queue">long (max runtime 2 weeks, 64 GB RAM)</option>
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104 <option value="extra_long_slow_queue">extra long (max runtime 4 weeks, 64 GB RAM)</option>
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105 </param>
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106 <when value="basic_fast_queue">
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107 <param name="queue_specification" type="text" label="Modify parameters (optional)"
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108 value="-l select=1:ncpus=10:mem=32gb:scratch_local=50gb -l walltime=48:00:00 -q elixirre@pbs.elixir-czech.cz -v TAREAN_MAX_MEM=4000000,TAREAN_CPU=4" />
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109 </when>
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110
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111 <when value="long_slow_queue">
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112 <param name="queue_specification" type="text" label="Modify parameters (optional)"
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113 value="-l select=1:ncpus=16:mem=112gb:scratch_local=50gb -l walltime=336:00:00 -q elixirre@pbs.elixir-czech.cz -v TAREAN_MAX_MEM=64000000,TAREAN_CPU=15" />
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114 </when>
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115 <when value="extra_long_slow_queue">
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116 <param name="queue_specification" type="text" label="Modify parameters (optional)"
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117 value="-l select=1:ncpus=16:mem=112gb:scratch_local=50gb -l walltime=720:00:00 -q elixirre@pbs.elixir-czech.cz -v TAREAN_MAX_MEM=64000000,TAREAN_CPU=15" />
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118 </when>
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119 </conditional>
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120
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121
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122
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123 </inputs>
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124 <outputs>
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125 <data name="log" format="txt" label="TAREAN log file"/>
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126 <data name="ReportArchive" format="zip" label="TAREAN Archive with HTML report from data ${FastaFile.hid}"/>
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127 <data name="ReportFile" format="html" label="TAREAN HTML report from data ${FastaFile.hid}"/>
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128 </outputs>
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129
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130 <help>
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131 **HELP**
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132
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133 TAREAN - TAndem REpeat ANalyzer is a computational pipeline for
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134 **unsupervised identification of satellite repeats** from unassembled
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135 sequence reads. The pipeline uses low-pass paired-end whole genome
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136 sequence reads and performs graph-based clustering. The resulting
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137 clusters, representing all types of repeats present in the genome, are
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138 then examined to identify those containing circular structures indicative
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139 of tandem repeats. A poster summarizing TAREAN principles and
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140 implementation can be found `here.`__
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141
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142
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143 .. __: http://w3lamc.umbr.cas.cz/lamc/?page_id=312
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144
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145 **Input data**
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146
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147
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148 The analysis requires **paired-end reads** generated by whole genome
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149 shotgun sequencing. The data should be provided as a single input file in
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150 fasta format with the reads interlaced (see example below). All the pairs
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151 must be complete, i.e. both "forward" and "reverse" sequence reads must be
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152 present. The reads should all be trimmed to the same length. The optimal
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153 size range is between 100 and 200 nucleotides. The number of reads to be
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154 analyzed should not exceed 1x coverage of the genome. Genome coverage
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155 between 0.01 and 0.5x is recommended. The reads should be filtered for
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156 quality. The recommended quality filtering is as follows: each read should
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157 have a quality score >=10 for 95% of the bases, i.e. if your reads are 100
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158 base pairs long, then a read only passes this quality threshold if 95
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159 bases have a quality of 10 or higher. Additionally, any reads containing
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160 indeterminate base pairs (indicated as N in the reads) should be removed.
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161 Finally, if either one of the reads in a pair fails to meet the
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162 aforementioned thresholds, **both** sequences should be removed.
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163 example of interlaced input format::
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164
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165 >0001_f
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166 CGTAATATACATACTTGCTAGCTAGTTGGATGCATCCAACTTGCAAGCTAGTTTGATG
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167 >0001_r
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168 GATTTGACGGACACACTAACTAGCTAGTTGCATCTAAGCGGGCACACTAACTAACTAT
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169 >0002_f
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170 ACTCATTTGGACTTAACTTTGATAATAAAAACTTAAAAAGGTTTCTGCACATGAATCG
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171 >0002_r
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172 TATGTTGAAAAATTGAATTTCGGGACGAAACAGCGTCTATCGTCACGACATAGTGCTC
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173 >0003_f
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174 TGACATTTGTGAACGTTAATGTTCAACAAATCTTTCCAATGTCTTTTTATCTTATCAT
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175 >0003_r
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176 TATTGAAATACTGGACACAAATTGGAAATGAAACCTTGTGAGTTATTCAATTTATGTT
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177 ...
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178
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179
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180 To perform the quality filtering on your fastQ formatted data as described
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181 above, and to interlace your paired-end sequence reads,
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182 please use the `Preprocessing of paired-reads`__ tool.
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183
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184 .. __: tool_runner?tool_id=paired_fastq_filtering
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185
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186
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187 **Additional parameters**
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188
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189 **Sample size** defines how many reads will be used during the computation.
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190 The default setting of 500,000 reads will enable detection of high copy
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191 number satellites within several hours. For higher
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192 sensitivity the sample size can be increased. Since the sample size affects
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193 memory usage, this parameter may be automatically adjusted to a lower value
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194 during the run. The maximum sample size which can be processed depends on the
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195 repetitiveness of the analyzed genome. This significantly limits the number of reads
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196 that can be analyzed with the TAREAN pipeline.
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197
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198 **Perform cluster merging**. Families of repetitive elements are
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199 frequently split into multiple clusters rather than being represented as a
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200 single one. If you do not want to merge clusters based on the presence
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201 of broken read pairs, disable this option.
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202
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203 **Use custom repeat database**. This option allows users to perform similarity
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204 comparison of identified repeats to their custom databases. The repeat class should
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205 be encoded in FASTA headers of database entries in order to allow correct
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206 parsing of similarity hits.
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207
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208 **Similarity search options** By default sequence reads are compared using
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209 mgblast program. Default threshold is explicitly set to 90% sequence
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210 similarity spanning at least 55% of the read length (in the case of reads
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211 differing in length it applies to the longer one). Additionally, sequence
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212 overlap must be at least 55 nt. If you select option for shorter reads
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213 than 100 nt, minimum overlap 55 nt is not required.
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214
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215 By default,
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216 mgblast search use DUST program to filter out
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217 low-complexity sequences. If you want
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218 to increase sensitivity of detection of satellites with shorter monomer
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219 use option with '*no masking of low complexity repeats*'. Note that omitting
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220 DUST filtering will significantly increase running times
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221
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222 **Output**
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223
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224 A list of clusters identified as putative satellite repeats, their genomic
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225 abundance and various cluster characteristics are provided. Length and
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226 consensus sequences of reconstructed monomers are also shown and
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227 accompanied by a detailed output from kmer-based reconstruction including
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228 sequences and sequence logos of alternative variants of monomer sequences.
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229
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230 The output includes an **HTML summary** with a table listing all analyzed
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231 clusters. More detailed information about clusters is provided in
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232 additional files and directories. All results are also provided as a
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233 downloadable **zip archive**. Since read clustering results in
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234 thousands of clusters, the search for satellite repeats is limited to
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235 a subset of the largest ones corresponding to the most abundant genomic
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236 repeats. The default setting of the pipeline is to analyze all clusters containing at least
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237 0.01% of the input reads. Besides the satellite repeats, three other
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238 groups of clusters are reported in the output (1) LTR-retrotransposons,
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239 (2) 45S and 5S rDNA and (3) all remaining clusters passing the size
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240 threshold. As (1) and (2) contain sequences with circular
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241 graphs, their consensus is calculated in the same way as for satellite
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242 repeats. Additionally a **log file** reporting the progress of the
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243 computational pipeline is provided.
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244
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245
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246 </help>
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247
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248 </tool>
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