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19
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1 <tool id="tarean" name="Tandem Repeat Analyzer" version="2.3.9.1" >
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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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19
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10 <container type="singularity">library://repeatexplorer/default/repex_tarean:0.3.9-579a65d</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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16
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14 export TAREAN_CPU=\$GALAXY_SLOTS;
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19
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15 export TAREAN_MAX_MEM=\$((GALAXY_MEMORY_MB*1024));
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4
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16 seqclust --paired --sample ${read_sampling.sample} --output_dir=tarean_output --logfile=${log} --cleanup --tarean_mode
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2
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17 #if $advanced_options.advanced:
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18 --mincl $advanced_options.size_threshold $advanced_options.keep_names $advanced_options.automatic_filtering -M $advanced_options.merging
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19 #if $advanced_options.custom_library.options_custom_library :
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20 -d $advanced_options.custom_library.library extra_database
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21 #end if
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22 #if $advanced_options.options.options:
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23 -opt $advanced_options.options.options
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24 #end if
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25 #else:
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26 -M 0.2
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27
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28 #end if
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29 ${FastaFile} >stdout.log 2> stderr.log ;
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30 echo "STDOUT CONTENT:" >> ${log} ;
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31 cat stdout.log >> ${log} ;
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32 echo "STDERR CONTENT:" >> ${log} ;
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33 cat stderr.log >> ${log} &&
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34 /opt/repex_tarean/stderr_filter.py stderr.log &&
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2
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35 cd tarean_output &&
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36 zip -r ${ReportArchive}.zip * &&
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37 mv ${ReportArchive}.zip ${ReportArchive} &&
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38 cp index.html ${ReportFile} &&
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39 mkdir -p ${ReportFile.extra_files_path} &&
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40 cp -r --parents libdir ${ReportFile.extra_files_path} &&
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41 cp -r --parents seqclust/clustering/superclusters ${ReportFile.extra_files_path} &&
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42 cp -r --parents seqclust/clustering/clusters ${ReportFile.extra_files_path} &&
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43 cp seqclust/clustering/hitsort.cls ${ReportFile.extra_files_path}/seqclust/clustering/hitsort.cls &&
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44 cp *.png ${ReportFile.extra_files_path}/ &&
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45 cp *.csv ${ReportFile.extra_files_path}/ &&
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46 cp *.html ${ReportFile.extra_files_path}/ &&
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47 cp *.css ${ReportFile.extra_files_path}/ &&
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48 cp *.fasta ${ReportFile.extra_files_path}/ 2>>$log && rm -r ../tarean_output || :
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2
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49
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50
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51 </command>
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52
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53 <inputs>
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54 <param name="FastaFile" label="Paired-end Illumina reads" type="data" format="fasta"
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55 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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56
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57 <conditional name="read_sampling">
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58 <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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59 <when value="false">
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60 <!-- pass -->
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16
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61 <param name="sample" label="Sample size" type="hidden" value="0" help="Number of analyzed reads"/>
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2
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62 </when>
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63 <when value="true">
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64 <param name="sample" label="Sample size" type="integer" value="500000" min="10000" help="Number of analyzed reads"/>
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65 </when>
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66 </conditional>
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67
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68 <conditional name="advanced_options">
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69 <param name="advanced" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Advanced options" />
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70 <when value="false">
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71 <!-- pass -->
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72 </when>
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73 <when value="true">
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74 <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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75 <conditional name="custom_library">
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76 <param name="options_custom_library" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Use custom repeat database"/>
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77 <when value="false">
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78 <!-- do nothing here -->
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79 </when>
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80 <when value="true">
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81 <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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82 </when>
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83 </conditional>
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84 <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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85 <param name="automatic_filtering" label="Perform automatic filtering of abundant satellite repeats" type="boolean" truevalue="--automatic_filtering" falsevalue="" checked="false"/>
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86 <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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87 <conditional name="options">
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88 <param name="options" type="select" label="Similarity search options">
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89 <option value="ILLUMINA" selected="true">Default </option>
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90 <option value="ILLUMINA_DUST_OFF" selected="false">Masking of low complexity repeats disabled </option>
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91
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92 <!-- <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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93 <!-- <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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94 <!-- <option value="OXFORD_NANOPORE" selected="false"> -->
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95 <!-- Pseudo short reads simulated from Oxford Nanopore data, experimental feature! -->
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96 <!-- </option> -->
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97 </param>
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98 </conditional>
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99 </when>
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100 </conditional>
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101
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102 <param name="queue_select" type="select" label="Select queue">
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103 <option value="basic_fast_queue">basic (max runtime 2 days, 4 GB RAM)</option>
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104 <option value="long_slow_queue">long (max runtime 2 weeks, 64 GB RAM)</option>
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105 <option value="extra_long_slow_queue">extra long (max runtime 4 weeks, 64 GB RAM)</option>
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106 </param>
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2
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107
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108 </inputs>
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109 <outputs>
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110 <data name="log" format="txt" label="TAREAN log file"/>
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111 <data name="ReportArchive" format="zip" label="TAREAN Archive with HTML report from data ${FastaFile.hid}"/>
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112 <data name="ReportFile" format="html" label="TAREAN HTML report from data ${FastaFile.hid}"/>
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113 </outputs>
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114
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115 <help>
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116 **HELP**
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117
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118 TAREAN - TAndem REpeat ANalyzer is a computational pipeline for
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119 **unsupervised identification of satellite repeats** from unassembled
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120 sequence reads. The pipeline uses low-pass paired-end whole genome
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121 sequence reads and performs graph-based clustering. The resulting
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122 clusters, representing all types of repeats present in the genome, are
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123 then examined to identify those containing circular structures indicative
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124 of tandem repeats. A poster summarizing TAREAN principles and
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125 implementation can be found `here.`__
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126
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127
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128 .. __: http://w3lamc.umbr.cas.cz/lamc/?page_id=312
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129
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130 **Input data**
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131
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132
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133 The analysis requires **paired-end reads** generated by whole genome
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134 shotgun sequencing. The data should be provided as a single input file in
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135 fasta format with the reads interlaced (see example below). All the pairs
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136 must be complete, i.e. both "forward" and "reverse" sequence reads must be
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137 present. The reads should all be trimmed to the same length. The optimal
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138 size range is between 100 and 200 nucleotides. The number of reads to be
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139 analyzed should not exceed 1x coverage of the genome. Genome coverage
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140 between 0.01 and 0.5x is recommended. The reads should be filtered for
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141 quality. The recommended quality filtering is as follows: each read should
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142 have a quality score >=10 for 95% of the bases, i.e. if your reads are 100
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143 base pairs long, then a read only passes this quality threshold if 95
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144 bases have a quality of 10 or higher. Additionally, any reads containing
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145 indeterminate base pairs (indicated as N in the reads) should be removed.
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146 Finally, if either one of the reads in a pair fails to meet the
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147 aforementioned thresholds, **both** sequences should be removed.
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148 example of interlaced input format::
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149
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150 >0001_f
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151 CGTAATATACATACTTGCTAGCTAGTTGGATGCATCCAACTTGCAAGCTAGTTTGATG
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152 >0001_r
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153 GATTTGACGGACACACTAACTAGCTAGTTGCATCTAAGCGGGCACACTAACTAACTAT
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154 >0002_f
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155 ACTCATTTGGACTTAACTTTGATAATAAAAACTTAAAAAGGTTTCTGCACATGAATCG
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156 >0002_r
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157 TATGTTGAAAAATTGAATTTCGGGACGAAACAGCGTCTATCGTCACGACATAGTGCTC
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158 >0003_f
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159 TGACATTTGTGAACGTTAATGTTCAACAAATCTTTCCAATGTCTTTTTATCTTATCAT
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160 >0003_r
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161 TATTGAAATACTGGACACAAATTGGAAATGAAACCTTGTGAGTTATTCAATTTATGTT
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162 ...
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163
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164
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165 To perform the quality filtering on your fastQ formatted data as described
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166 above, and to interlace your paired-end sequence reads,
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167 please use the `Preprocessing of paired-reads`__ tool.
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168
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169 .. __: tool_runner?tool_id=paired_fastq_filtering
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170
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171
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172 **Additional parameters**
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173
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174 **Sample size** defines how many reads will be used during the computation.
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175 The default setting of 500,000 reads will enable detection of high copy
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176 number satellites within several hours. For higher
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177 sensitivity the sample size can be increased. Since the sample size affects
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178 memory usage, this parameter may be automatically adjusted to a lower value
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179 during the run. The maximum sample size which can be processed depends on the
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180 repetitiveness of the analyzed genome. This significantly limits the number of reads
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181 that can be analyzed with the TAREAN pipeline.
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182
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183 **Perform cluster merging**. Families of repetitive elements are
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184 frequently split into multiple clusters rather than being represented as a
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185 single one. If you do not want to merge clusters based on the presence
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186 of broken read pairs, disable this option.
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187
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188 **Use custom repeat database**. This option allows users to perform similarity
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189 comparison of identified repeats to their custom databases. The repeat class should
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190 be encoded in FASTA headers of database entries in order to allow correct
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191 parsing of similarity hits.
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192
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193 **Similarity search options** By default sequence reads are compared using
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194 mgblast program. Default threshold is explicitly set to 90% sequence
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195 similarity spanning at least 55% of the read length (in the case of reads
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196 differing in length it applies to the longer one). Additionally, sequence
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197 overlap must be at least 55 nt. If you select option for shorter reads
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198 than 100 nt, minimum overlap 55 nt is not required.
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199
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200 By default,
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201 mgblast search use DUST program to filter out
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202 low-complexity sequences. If you want
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203 to increase sensitivity of detection of satellites with shorter monomer
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204 use option with '*no masking of low complexity repeats*'. Note that omitting
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205 DUST filtering will significantly increase running times
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206
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207 **Output**
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208
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209 A list of clusters identified as putative satellite repeats, their genomic
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210 abundance and various cluster characteristics are provided. Length and
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211 consensus sequences of reconstructed monomers are also shown and
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212 accompanied by a detailed output from kmer-based reconstruction including
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213 sequences and sequence logos of alternative variants of monomer sequences.
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214
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215 The output includes an **HTML summary** with a table listing all analyzed
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216 clusters. More detailed information about clusters is provided in
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217 additional files and directories. All results are also provided as a
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218 downloadable **zip archive**. Since read clustering results in
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219 thousands of clusters, the search for satellite repeats is limited to
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220 a subset of the largest ones corresponding to the most abundant genomic
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221 repeats. The default setting of the pipeline is to analyze all clusters containing at least
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222 0.01% of the input reads. Besides the satellite repeats, three other
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223 groups of clusters are reported in the output (1) LTR-retrotransposons,
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224 (2) 45S and 5S rDNA and (3) all remaining clusters passing the size
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225 threshold. As (1) and (2) contain sequences with circular
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226 graphs, their consensus is calculated in the same way as for satellite
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227 repeats. Additionally a **log file** reporting the progress of the
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228 computational pipeline is provided.
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229
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230
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231 </help>
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232
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233 </tool>
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