annotate bowtie2_wrapper.xml @ 8:82414e16b6bd draft

planemo upload for repository https://github.com/galaxyproject/tools-devteam/tree/master/tools/bowtie2 commit 637e614b544bf3cd75306f27bcfe537cacae4a58
author devteam
date Mon, 16 Nov 2015 14:38:37 -0500
parents 4f92dccc808a
children a9d4f71dbfb0
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1 <tool id="bowtie2" name="Bowtie2" version="2.2.6">
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2 <description>- map reads against reference genome</description>
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3 <macros>
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4 <import>read_group_macros.xml</import>
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5 </macros>
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6 <version_command>bowtie2 --version</version_command>
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7 <requirements>
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8 <requirement type="package" version="2.2.6">bowtie2</requirement>
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9 <requirement type="package" version="1.2">samtools</requirement>
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10 </requirements>
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11 <command>
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12
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13 ## prepare bowtie2 index
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14 #set index_path = ''
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15 #if str($reference_genome.source) == "history":
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16 bowtie2-build "$reference_genome.own_file" genome &amp;&amp;
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17 ln -s "$reference_genome.own_file" genome.fa &amp;&amp;
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18 #set index_path = 'genome'
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19 #else:
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20 #set index_path = $reference_genome.index.fields.path
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21 #end if
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22
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23 ## execute bowtie2
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24
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25 bowtie2
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26
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27 ## number of threads
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28 -p \${GALAXY_SLOTS:-4}
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29
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30 ## index file path
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31 -x $index_path
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32
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33 ## Fastq inputs
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34 #if str( $library.type ) == "single":
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35 -U "${library.input_1}"
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36 #if str( $library.unaligned_file ) == "true":
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37 --un $output_unaligned_reads_l
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38 #end if
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39 #elif str( $library.type ) == "paired":
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40 -1 "${library.input_1}"
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41 -2 "${library.input_2}"
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42 #if str( $library.paired_options.paired_options_selector ) == "yes":
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43 -I "${library.paired_options.I}"
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44 -X "${library.paired_options.X}"
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45 ${library.paired_options.fr_rf_ff}
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46 ${library.paired_options.no_mixed}
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47 ${library.paired_options.no_discordant}
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48 ${library.paired_options.dovetail}
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49 ${library.paired_options.no_contain}
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50 ${library.paired_options.no_overlap}
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51 #end if
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52 #if str( $library.unaligned_file ) == "true":
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53 --un-conc $output_unaligned_reads_l
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54 #end if
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55 #else
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56 ## prepare collection
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57 -1 $library.input_1.forward
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58 -2 $library.input_1.reverse
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59 #if str( $library.paired_options.paired_options_selector ) == "yes":
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60 -I "${library.paired_options.I}"
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61 -X "${library.paired_options.X}"
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62 ${library.paired_options.fr_rf_ff}
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63 ${library.paired_options.no_mixed}
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64 ${library.paired_options.no_discordant}
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65 ${library.paired_options.dovetail}
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66 ${library.paired_options.no_contain}
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67 ${library.paired_options.no_overlap}
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68 #end if
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69 #if str( $library.unaligned_file ) == "true":
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70 --un-conc $output_unaligned_reads_l
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71 #end if
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72 #end if
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73
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74 ## Read group information.
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75 @define_read_group_helpers@
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76 #if str( $library.type ) == "single":
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77 #set $rg_auto_name = $read_group_name_default($library.input_1)
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78 #elif str( $library.type ) == "paired":
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79 #set $rg_auto_name = $read_group_name_default($library.input_1, $library.input_2)
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80 #else
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81 #set $rg_auto_name = $read_group_name_default($library.input_1)
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82 #end if
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83 @set_use_rg_var@
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84 @set_read_group_vars@
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85 #if $use_rg
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86 $format_read_group("", $rg_id, '"', arg='--rg-id ')
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87 $format_read_group("SM:", $rg_sm, '"', arg='--rg ')
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88 $format_read_group("PL:", $rg_pl, '"', arg='--rg ')
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89 $format_read_group("LB:", $rg_lb, '"', arg='--rg ')
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90 $format_read_group("CN:", $rg_cn, '"', arg='--rg ')
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91 $format_read_group("DS:", $rg_ds, '"', arg='--rg ')
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92 $format_read_group("DT:", $rg_dt, '"', arg='--rg ')
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93 $format_read_group("FO:", $rg_fo, '"', arg='--rg ')
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94 $format_read_group("KS:", $rg_ks, '"', arg='--rg ')
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95 $format_read_group("PG:", $rg_pg, '"', arg='--rg ')
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96 $format_read_group("PI:", $rg_pi, '"', arg='--rg ')
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97 $format_read_group("PU:", $rg_pu, '"', arg='--rg ')
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98 #end if
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99
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100 ## Analysis type
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101 #if ( str( $analysis_type.analysis_type_selector ) == "simple" and str( $analysis_type.presets ) != "no_presets" ):
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102 $analysis_type.presets
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103 #elif str( $analysis_type.analysis_type_selector ) == "full":
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104 #if str( $analysis_type.input_options.input_options_selector ) == "yes":
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105 --skip "${analysis_type.input_options.skip}"
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106 --qupto "${analysis_type.input_options.qupto}"
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107 --trim5 "${analysis_type.input_options.trim5}"
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108 --trim3 "${analysis_type.input_options.trim3}"
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109 ${analysis_type.input_options.qv_encoding}
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110 ${analysis_type.input_options.solexa_quals}
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111 ${analysis_type.input_options.int_quals}
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112 #end if
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113
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114 #if str( $analysis_type.alignment_options.alignment_options_selector ) == "yes":
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115 -N "${analysis_type.alignment_options.N}"
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116 -L "${analysis_type.alignment_options.L}"
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117 -i "${analysis_type.alignment_options.i}"
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118 --n-ceil "${analysis_type.alignment_options.n_ceil}"
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119 --dpad "${analysis_type.alignment_options.dpad}"
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120 --gbar "${analysis_type.alignment_options.gbar}"
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121 ${analysis_type.alignment_options.ignore_quals}
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122 ${analysis_type.alignment_options.nofw}
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123 ${analysis_type.alignment_options.norc}
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124 ${analysis_type.alignment_options.no_1mm_upfront}
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125 #if str( $analysis_type.alignment_options.align_mode.align_mode_selector ) == "end-to-end":
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126 --end-to-end
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127 --score-min "${analysis_type.alignment_options.align_mode.score_min_ete}"
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128 #elif str( $analysis_type.alignment_options.align_mode.align_mode_selector ) == "local":
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129 --local
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130 --score-min "${analysis_type.alignment_options.align_mode.score_min_loc}"
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131 #end if
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132 #end if
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133
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134 #if str( $analysis_type.scoring_options.scoring_options_selector ) == "yes":
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135 #if ( str( $analysis_type.alignment_options.alignment_options_selector ) == "yes" and str( $analysis_type.alignment_options.align_mode.align_mode_selector ) == "local" ):
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136 --ma "${analysis_type.scoring_options.ma}"
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137 #end if
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138 --mp "${analysis_type.scoring_options.mp}"
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139 --np "${analysis_type.scoring_options.np}"
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140 --rdg "${analysis_type.scoring_options.rdg_read_open},${analysis_type.scoring_options.rdg_read_extend}"
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141 --rfg "${analysis_type.scoring_options.rfg_ref_open},${analysis_type.scoring_options.rfg_ref_extend}"
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142 #end if
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143
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144 #if str( $analysis_type.reporting_options.reporting_options_selector ) == "k":
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145 -k "${analysis_type.reporting_options.k}"
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146 #elif str( $analysis_type.reporting_options.reporting_options_selector ) == "a":
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147 -a
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148 #end if
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149
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150 #if str( $analysis_type.effort_options.effort_options_selector ) == "yes":
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151 -D "${analysis_type.effort_options.D}"
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152 -R "${analysis_type.effort_options.R}"
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153 #end if
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154
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155 #if str( $analysis_type.sam_options.sam_options_selector ) == "yes":
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156 ${analysis_type.sam_options.no_unal}
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157 ${analysis_type.sam_options.omit_sec_seq}
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158 #end if
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159
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160 #if str( $analysis_type.other_options.other_options_selector ) == "yes":
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161 ${analysis_type.other_options.reorder}
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162 ${analysis_type.other_options.non_deterministic}
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163 --seed "${analysis_type.other_options.seed}"
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164 #end if
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165
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166 #elif str( $analysis_type.analysis_type_selector ) == "cline":
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167 ${analysis_type.cline}
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168 #end if
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169
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170 ## output file
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171 #if ( str( $analysis_type.analysis_type_selector ) != "full" or str( $analysis_type.sam_opt ) != "true" ):
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172 | samtools view -Su - | samtools sort -o - - &gt; $output
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173 #else
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174 &gt; $output_sam
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175 #end if
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176
0
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177 ## rename unaligned sequence files
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178 #if $library.type == "paired" and $output_unaligned_reads_l and $output_unaligned_reads_r:
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179 #from os.path import splitext
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180 #set _unaligned_root, _unaligned_ext = splitext( str( $output_unaligned_reads_l ) )
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181 &amp;&amp; mv "${ _unaligned_root }.1${_unaligned_ext}" "${ output_unaligned_reads_l }"
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182 &amp;&amp; mv "${ _unaligned_root }.2${_unaligned_ext}" "${ output_unaligned_reads_r }"
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183 #end if
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184
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185 </command>
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186 <!-- basic error handling -->
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187 <stdio>
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188 <exit_code range="1:" level="fatal" description="Tool exception" />
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189 </stdio>
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190
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191 <inputs>
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192 <!-- single/paired -->
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193 <conditional name="library">
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194 <param name="type" type="select" label="Is this single or paired library">
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195 <option value="single">Single-end</option>
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196 <option value="paired">Paired-end</option>
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197 <option value="paired_collection">Paired-end Dataset Collection</option>
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198 </param>
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199
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200 <when value="single">
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201 <param name="input_1" format="fastqsanger" type="data" label="FASTQ file" help="Must be of datatype &quot;fastqsanger&quot;" />
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202 <param name="unaligned_file" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Write unaligned reads (in fastq format) to separate file(s)" help="--un/--un-conc; This triggers --un parameter for single reads and --un-conc for paired reads" />
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203 </when>
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204 <when value="paired">
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205 <param name="input_1" format="fastqsanger" type="data" label="FASTQ file #1" help="Must be of datatype &quot;fastqsanger&quot;" />
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206 <param name="input_2" format="fastqsanger" type="data" label="FASTQ file #2" help="Must be of datatype &quot;fastqsanger&quot;" />
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207 <param name="unaligned_file" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Write unaligned reads (in fastq format) to separate file(s)" help="--un/--un-conc; This triggers --un parameter for single reads and --un-conc for paired reads" />
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208 <conditional name="paired_options">
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209 <param name="paired_options_selector" type="select" label="Do you want to set paired-end options?" help="See &quot;Alignment Options&quot; section of Help below for information">
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210 <option value="no" selected="True">No</option>
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211 <option value="yes">Yes</option>
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212 </param>
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213 <when value="yes">
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214 <param name="I" type="integer" value="0" min="0" label="Set the minimum fragment length for valid paired-end alignments" help="-I/--minins; E.g. if `-I 60` is specified and a paired-end alignment consists of two 20-bp alignments in the appropriate orientation with a 20-bp gap between them, that alignment is considered valid (as long as `-X` is also satisfied). A 19-bp gap would not be valid in that case. If trimming options `-3` or `-5` are also used, the `-I` constraint is applied with respect to the untrimmed mates. The larger the difference between `-I` and `-X`, the slower Bowtie 2 will run. This is because larger differences bewteen `-I` and `-X` require that Bowtie 2 scan a larger window to determine if a concordant alignment exists. For typical fragment length ranges (200 to 400 nucleotides), Bowtie 2 is very efficient. Default=0"/>
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215 <param name="X" type="integer" value="500" min="0" label="Set the maximum fragment length for valid paired-end alignments" help="-X/--maxins; E.g. if `-X 100` is specified and a paired-end alignment consists of two 20-bp alignments in the proper orientation with a 60-bp gap between them, that alignment is considered valid (as long as `-I` is also satisfied). A 61-bp gap would not be valid in that case. If trimming options `-3` or `-5` are also used, the `-X` constraint is applied with respect to the untrimmed mates, not the trimmed mates; Default=500"/>
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216 <param name="fr_rf_ff" type="select" display="radio" label="Select the upstream/downstream mate orientations for a valid paired-end alignment against the forward reference strand" help="--fr, --rf, or --ff; E.g., if `--fr` is specified and there is a candidate paired-end alignment where mate 1 appears upstream of the reverse complement of mate 2 and the fragment length constraints (`-I` and `-X`) are met, that alignment is valid. Also, if mate 2 appears upstream of the reverse complement of mate 1 and all other constraints are met, that too is valid. `--rf` likewise requires that an upstream mate1 be reverse-complemented and a downstream mate2 be forward-oriented. `--ff` requires both an upstream mate 1 and a downstream mate 2 to be forward-oriented; Default=--fr (appropriate for Illumina's Paired-end Sequencing Assay)">
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217 <option value="--fr" selected="True">--fr</option>
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218 <option value="--rf">--rf</option>
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219 <option value="--ff">--ff</option>
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220 </param>
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221 <param name="no_mixed" type="boolean" truevalue="--no-mixed" falsevalue="" checked="False" label="Disable no-mixed behavior" help="--no-mixed; By default, when `bowtie2` cannot find a concordant or discordant alignment for a pair, it then tries to find alignments for the individual mates; default=False"/>
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222 <param name="no_discordant" type="boolean" truevalue="--no-discordant" falsevalue="" checked="False" label="Disable no-discordant behavior" help="--no-discordant; By default, `bowtie2` looks for discordant alignments if it cannot find any concordant alignments. A discordant alignment is an alignment where both mates align uniquely, but that does not satisfy the paired-end constraints (`--fr`/`--rf`/`--ff`, `-I`, `-X`); default=False"/>
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223 <param name="dovetail" type="boolean" truevalue="--dovetail" falsevalue="" checked="False" label="Allow mate dovetailing" help="--dovetail; If the mates `dovetail`, that is if one mate alignment extends past the beginning of the other such that the wrong mate begins upstream, consider that to be concordant. Default=False"/>
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224 <param name="no_contain" type="boolean" truevalue="--no-contain" falsevalue="" checked="False" label="Disallow one mate alignment to contain another" help="--no-contain; If one mate alignment contains the other, consider that to be non-concordant. Default=False"/>
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225 <param name="no_overlap" type="boolean" truevalue="--no-overlap" falsevalue="" checked="False" label="Disallow mate alignments to overlap" help="--no-overlap; If one mate alignment overlaps the other at all, consider that to be non-concordant. Default=False"/>
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226 </when>
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227 <when value="no">
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228 <!-- do nothing -->
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229 </when>
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230 </conditional>
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231 </when>
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232 <when value="paired_collection">
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233 <param name="input_1" format="fastqsanger" type="data_collection" collection_type="paired" label="FASTQ Paired Dataset" help="Must be of datatype &quot;fastqsanger&quot;" />
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234 <param name="unaligned_file" type="boolean" truevalue="true" falsevalue="false" checked="False" label="Write unaligned reads (in fastq format) to separate file(s)" help="--un/--un-conc; This triggers --un parameter for single reads and --un-conc for paired reads" />
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235 <conditional name="paired_options">
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236 <param name="paired_options_selector" type="select" label="Do you want to set paired-end options?" help="See &quot;Alignment Options&quot; section of Help below for information">
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237 <option value="no" selected="True">No</option>
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238 <option value="yes">Yes</option>
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239 </param>
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240 <when value="yes">
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241 <param name="I" type="integer" value="0" min="0" label="Set the minimum fragment length for valid paired-end alignments" help="-I/--minins; E.g. if `-I 60` is specified and a paired-end alignment consists of two 20-bp alignments in the appropriate orientation with a 20-bp gap between them, that alignment is considered valid (as long as `-X` is also satisfied). A 19-bp gap would not be valid in that case. If trimming options `-3` or `-5` are also used, the `-I` constraint is applied with respect to the untrimmed mates. The larger the difference between `-I` and `-X`, the slower Bowtie 2 will run. This is because larger differences bewteen `-I` and `-X` require that Bowtie 2 scan a larger window to determine if a concordant alignment exists. For typical fragment length ranges (200 to 400 nucleotides), Bowtie 2 is very efficient. Default=0"/>
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242 <param name="X" type="integer" value="500" min="0" label="Set the maximum fragment length for valid paired-end alignments" help="-X/--maxins; E.g. if `-X 100` is specified and a paired-end alignment consists of two 20-bp alignments in the proper orientation with a 60-bp gap between them, that alignment is considered valid (as long as `-I` is also satisfied). A 61-bp gap would not be valid in that case. If trimming options `-3` or `-5` are also used, the `-X` constraint is applied with respect to the untrimmed mates, not the trimmed mates; Default=500"/>
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243 <param name="fr_rf_ff" type="select" display="radio" label="Select the upstream/downstream mate orientations for a valid paired-end alignment against the forward reference strand" help="--fr, --rf, or --ff; E.g., if `--fr` is specified and there is a candidate paired-end alignment where mate 1 appears upstream of the reverse complement of mate 2 and the fragment length constraints (`-I` and `-X`) are met, that alignment is valid. Also, if mate 2 appears upstream of the reverse complement of mate 1 and all other constraints are met, that too is valid. `--rf` likewise requires that an upstream mate1 be reverse-complemented and a downstream mate2 be forward-oriented. `--ff` requires both an upstream mate 1 and a downstream mate 2 to be forward-oriented; Default=--fr (appropriate for Illumina's Paired-end Sequencing Assay)">
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244 <option value="--fr" selected="True">--fr</option>
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245 <option value="--rf">--rf</option>
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246 <option value="--ff">--ff</option>
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247 </param>
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248 <param name="no_mixed" type="boolean" truevalue="--no-mixed" falsevalue="" checked="False" label="Disable no-mixed behavior" help="--no-mixed; By default, when `bowtie2` cannot find a concordant or discordant alignment for a pair, it then tries to find alignments for the individual mates; default=False"/>
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249 <param name="no_discordant" type="boolean" truevalue="--no-discordant" falsevalue="" checked="False" label="Disable no-discordant behavior" help="--no-discordant; By default, `bowtie2` looks for discordant alignments if it cannot find any concordant alignments. A discordant alignment is an alignment where both mates align uniquely, but that does not satisfy the paired-end constraints (`--fr`/`--rf`/`--ff`, `-I`, `-X`); default=False"/>
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250 <param name="dovetail" type="boolean" truevalue="--dovetail" falsevalue="" checked="False" label="Allow mate dovetailing" help="--dovetail; If the mates `dovetail`, that is if one mate alignment extends past the beginning of the other such that the wrong mate begins upstream, consider that to be concordant. Default=False"/>
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251 <param name="no_contain" type="boolean" truevalue="--no-contain" falsevalue="" checked="False" label="Disallow one mate alignment to contain another" help="--no-contain; If one mate alignment contains the other, consider that to be non-concordant. Default=False"/>
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252 <param name="no_overlap" type="boolean" truevalue="--no-overlap" falsevalue="" checked="False" label="Disallow mate alignments to overlap" help="--no-overlap; If one mate alignment overlaps the other at all, consider that to be non-concordant. Default=False"/>
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253 </when>
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254 <when value="no">
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255 <!-- do nothing -->
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256 </when>
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257 </conditional>
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258 </when>
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259 </conditional>
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260
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261 <!-- reference genome -->
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262 <conditional name="reference_genome">
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263 <param name="source" type="select" label="Will you select a reference genome from your history or use a built-in index?" help="Built-ins were indexed using default options. See `Indexes` section of help below">
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264 <option value="indexed">Use a built-in genome index</option>
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265 <option value="history">Use a genome from the history and build index</option>
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266 </param>
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267 <when value="indexed">
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268 <param name="index" type="select" label="Select reference genome" help="If your genome of interest is not listed, contact the Galaxy team">
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269 <options from_data_table="bowtie2_indexes">
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270 <filter type="sort_by" column="2"/>
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271 <validator type="no_options" message="No indexes are available for the selected input dataset"/>
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272 </options>
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273 </param>
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274 </when>
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275 <when value="history">
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276 <param name="own_file" type="data" format="fasta" metadata_name="dbkey" label="Select reference genome" />
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277 </when>
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278 </conditional>
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279
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280 <!-- read group settings -->
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281 <expand macro="read_group_conditional" />
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282 <conditional name="analysis_type">
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283 <param name="analysis_type_selector" type="select" label="Select analysis mode">
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284 <option value="simple">1: Default setting only</option>
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285 <option value="full">2: Full parameter list</option>
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286 </param>
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287 <when value="simple">
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288 <param name="presets" type="select" display="radio" label="Do you want to use presets?" help="Allow selecting among several preset parameter settings. Choosing between these will result in dramatic changes in runtime. See help below to understand effects of these presets.">
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289 <option value="no_presets" selected="True">No, just use defaults</option>
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290 <option value="--very-fast">Very fast end-to-end (--very-fast)</option>
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291 <option value="--fast">Fast end-to-end (--fast)</option>
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292 <option value="--sensitive">Sensitive end-to-end (--sensitive)</option>
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293 <option value="--very-sensitive">Very sensitive end-to-end (--very-sensitive)</option>
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294 <option value="--very-fast-local">Very fast local (--very-fast-local)</option>
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295 <option value="--fast-local">Fast local (--fast-local)</option>
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296 <option value="--sensitive-local">Sensitive local (--sensitive-local)</option>
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297 <option value="--very-sensitive-local">Very sensitive local (--very-sensitive-local)</option>
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298 </param>
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299 </when>
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300 <when value="full">
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301 <conditional name="input_options">
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302 <param name="input_options_selector" type="select" label="Do you want to tweak input options?" help="See &quot;Input Options&quot; section of Help below for information">
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303 <option value="yes">Yes</option>
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304 <option value="no" selected="true">No</option>
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305 </param>
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306 <when value="yes">
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307 <param name="skip" type="integer" min="0" value="0" label="Skip (i.e. do not align) the first that many reads or pairs in the input" help="-s/--skip; default=0"/>
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308 <param name="qupto" type="integer" min="1" value="100000000" label="Align the first that many reads or read pairs from the input (after the -s/--skip reads or pairs have been skipped), then stop" help="-u/--qupto; for default behavior (no limit) leave this value very large"/>
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309 <param name="trim5" type="integer" min="0" value="0" label="Trim that many bases from 5' (left) end of each read before alignment" help="-5/--trim5; default=0"/>
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310 <param name="trim3" type="integer" min="0" value="0" label="Trim that many bases from 3' (right) end of each read before alignment" help="-3/--trim3; default=0"/>
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311 <param name="qv_encoding" type="select" display="radio" label="Select quality score encoding" help="See help below for more details">
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312 <option value="--phred33" selected="True">Input qualities are ASCII chars equal to the Phred quality plus 33. This is also called the "Phred+33" encoding, which is used by the very latest Illumina pipelines (--phred33)</option>
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313 <option value="--phred64">Input qualities are ASCII chars equal to the Phred quality plus 64. This is also called the "Phred+64" encoding (--phred64)</option>
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314 </param>
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315 <param name="solexa_quals" type="boolean" truevalue="--solexa-quals" falsevalue="" checked="False" label="Convert input qualities from Solexa (which can be negative) to Phred (which can't). This scheme was used in older Illumina GA Pipeline versions (prior to 1.3)" help="--solexa-quals; default=False"/>
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316 <param name="int_quals" type="boolean" truevalue="--int-quals" falsevalue="" checked="False" label="Quality values are represented in the read input file as space-separated ASCII integers, e.g., 40 40 30 40..., rather than ASCII characters, e.g., II?I.... Integers are treated as being on the Phred quality scale unless --solexa-quals is also specified" help="--int-quals; default=False"/>
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317 </when>
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318 <when value="no">
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319 <!-- do nothing -->
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320 </when>
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321 </conditional>
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322 <conditional name="alignment_options">
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323 <param name="alignment_options_selector" type="select" label="Do you want to tweak alignment options?" help="See &quot;Alignment Options&quot; section of Help below for information">
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324 <option value="yes">Yes</option>
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325 <option value="no" selected="true">No</option>
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326 </param>
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327 <when value="yes">
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328 <param name="N" type="integer" min="0" max="1" value="0" label="Set the number of mismatches to be allowed in a seed alignment during multiseed alignment (see `Multiseed alignment` section of help below)" help="-N; Can be set to 0 or 1. Setting this higher makes alignment slower (often much slower) but increases sensitivity; default=0"/>
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329 <param name="L" type="integer" min="0" max="32" value="22" label="Sets the length of the seed substrings to align during multiseed alignment (see `Multiseed alignment` section of help below)" help="-L; Smaller values make alignment slower but more sensitive. Default=22"/>
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330 <param name="i" type="text" value="S,1,1.15" label="Set a function governing the interval between seed substrings to use during multiseed alignment (see `Multiseed alignment` section of help below). Also see description of this option below in the help section" help="-i; Since it's best to use longer intervals for longer reads, this parameter sets the interval as a function of the read length, rather than a single one-size-fits-all number. For instance, specifying `-i S,1,2.5` sets the interval function `f` to `f(x) = 1 + 2.5 * sqrt(x)`, where x is the read length. If the function returns a result less than 1, it is rounded up to 1. Default=`S,1,1.15`"/>
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331 <param name="n_ceil" type="text" value="L,0,0.15" label="Set a function governing the maximum number of ambiguous characters (usually `N`s and/or `.`s) allowed in a read as a function of read length" help="--n-ceil; For instance, specifying `L,0,0.15` sets the N-ceiling function `f` to `f(x) = 0 + 0.15 * x`, where x is the read length. Reads exceeding this ceiling are filtered out. Default=`L,0,0.15`"/>
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332 <param name="dpad" type="integer" min="0" value="15" label="Pad dynamic programming problems by that many columns on either side to allow gaps" help="--dpad; default=15"/>
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333 <param name="gbar" type="integer" min="0" value="4" label="Disallow gaps within that many positions of the beginning or end of the read" help="--gbar; default=4"/>
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334 <param name="ignore_quals" type="boolean" truevalue="--ignore-quals" falsevalue="" selected="False" label="When calculating a mismatch penalty, always consider the quality value at the mismatched position to be the highest possible, regardless of the actual value" help="--ignore-quals; input is treated as though all quality values are high; default=False"/>
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335 <param name="nofw" type="boolean" truevalue="--nofw" falsevalue="" selected="False" label="Do not attempt to align unpaired reads to the forward (Watson) reference strand" help="In paired-end mode, `--nofw` and `--norc` pertain to the fragments; i.e. specifying `--nofw` causes `bowtie2` to explore only those paired-end configurations corresponding to fragments from the reverse-complement (Crick) strand. Default=False"/>
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336 <param name="norc" type="boolean" truevalue="--norc" falsevalue="" selected="False" label="Do not attempt to align unpaired reads to the reverse (Crick) reference strand" help="In paired-end mode, `--nofw` and `--norc` pertain to the fragments; i.e. specifying `--nofw` causes `bowtie2` to explore only those paired-end configurations corresponding to fragments from the reverse-complement (Crick) strand. Default=False"/>
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337 <param name="no_1mm_upfront" type="boolean" truevalue="--no-1mm-upfront" falsevalue="" selected="False" label="Prevent searching for 1-mismatch end-to-end alignments before using the multiseed heuristic (see `Multiseed alignment` section of help below)" help="--no-1mm-upfront; By default, Bowtie 2 will attempt to find either an exact or a 1-mismatch end-to-end alignment for the read *before* trying the multiseed heuristic. Such alignments can be found very quickly, and many short read alignments have exact or near-exact end-to-end alignments. However, this can lead to unexpected alignments when the user also sets options governing the multiseed heuristic, like `-L` and `-N`. For instance, if the user specifies `-N 0` and `-L` equal to the length of the read, the user will be surprised to find 1-mismatch alignments reported. This option prevents Bowtie 2 from searching for 1-mismatch end-to-end alignments before using the multiseed heuristic, which leads to the expected behavior when combined with options such as `-L` and `-N`. This comes at the expense of speed; Default=False"/>
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338 <conditional name="align_mode">
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339 <param name="align_mode_selector" type="select" display="radio" label="Select between `--local` and `--end-to-end` alignment modes" help="--local and --end-to-end; see help below for detailed explanation; default=--end-to-end">
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340 <option value="end-to-end" selected="True">End to End (--end-to-end)</option>
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341 <option value="local">Local (--local)</option>
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342 </param>
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343 <when value="end-to-end">
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344 <param name="score_min_ete" type="text" value="L,-0.6,-0.6" label="Set a function governing the minimum alignment score needed for an alignment to be considered `valid` (i.e. good enough to report)" help="--score-min; This is a function of read length. For instance, specifying `L,0,-0.6` sets the minimum-score function `f` to `f(x) = 0 + -0.6 * x`, where `x` is the read length. The default in `--end-to-end` mode is `L,-0.6,-0.6` and the default in `--local` mode is `G,20,8`"/>
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345 </when>
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346 <when value="local">
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347 <param name="score_min_loc" type="text" value="G,20,8" label="Set a function governing the minimum alignment score needed for an alignment to be considered `valid` (i.e. good enough to report)" help="--score-min; This is a function of read length. For instance, specifying `L,0,-0.6` sets the minimum-score function `f` to `f(x) = 0 + -0.6 * x`, where `x` is the read length. The default in `--end-to-end` mode is `L,-0.6,-0.6` and the default in `--local` mode is `G,20,8`"/>
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348 </when>
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349 </conditional>
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350 </when>
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351 <when value="no">
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352 <!-- do nothing -->
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353 </when>
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354 </conditional>
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355 <conditional name="scoring_options">
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356 <param name="scoring_options_selector" type="select" label="Do you want to tweak scoring options?" help="See &quot;Scoring Options&quot; section of Help below for information">
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357 <option value="yes">Yes</option>
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358 <option value="no" selected="true">No</option>
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359 </param>
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360 <when value="yes">
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361 <param name="ma" type="integer" value="2" label="Set the match bonus" help="--ma; In `--local` mode match bonus is added to the alignment score for each position where a read character aligns to a reference character and the characters match. Not used in `--end-to-end` mode; Default=2"/>
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362 <param name="mp" type="text" value="6,2" label="Set the maximum (`MX`) and minimum (`MN`) mismatch penalties, both integers" help="--mp; A number less than or equal to `MX` and greater than or equal to `MN` is subtracted from the alignment score for each position where a read character aligns to a reference character, the characters do not match, and neither is an `N`. If `--ignore-quals` is specified, the number subtracted quals `MX`. Otherwise, the number subtracted is `MN + floor( (MX-MN)(MIN(Q, 40.0)/40.0) )` where Q is the Phred quality value; Default=6,2"/>
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363 <param name="np" type="integer" value="1" label="Sets penalty for positions where the read, reference, or both, contain an ambiguous character such as `N`" help="--np; Default=1"/>
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364 <param name="rdg_read_open" type="integer" value="5" label="Set the read gap opening penalty" help="--rdg; this is the first component of --rdg flag - opening penalty; Default=5"/>
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365 <param name="rdg_read_extend" type="integer" value="3" label="Set the read gap extension penalty" help="--rdg; this is the second component of --rdg flag - extension penalty; Default=3"/>
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366 <param name="rfg_ref_open" type="integer" value="5" label="Set the reference gap opening penalty" help="--rfg; this is the first component of --rfg flag - opening penalty; Default=5"/>
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367 <param name="rfg_ref_extend" type="integer" value="3" label="Set the reference gap extension penalty" help="--rfg; this is the second component of --rfg flag - extension penalty; Default=3"/>
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368 </when>
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369 <when value="no">
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370 <!-- do nothing -->
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371 </when>
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372 </conditional>
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373 <conditional name="reporting_options">
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374 <param name="reporting_options_selector" type="select" label="Do you want to use -a or -k options" help="Make sure you understand implications of setting -k and -a. See &quot;Reporting Options&quot; section of Help below for information on -k and -a options">
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375 <option value="no" selected="true">No, do not set</option>
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376 <option value="k">Set -k option and enter -k value</option>
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377 <option value="a">Set -a option</option>
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378 </param>
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379 <when value="no">
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380 <!-- do nothing -->
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381 </when>
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382 <when value="k">
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383 <param name="k" type="integer" min="1" value="1" label="Searches for at most that many distinct, valid alignments for each read" help="-k; see detailed description of this option in the help section below. Note: Bowtie 2 is not designed with large values for `-k` in mind, and when aligning reads to long, repetitive genomes large `-k` can be very, very slow"/>
2
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384 </when>
3
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385 <when value="a">
2
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parents: 0
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386 <!-- do nothing here; set -a flag on the command line-->
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387 </when>
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388 </conditional>
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389 <conditional name="effort_options">
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390 <param name="effort_options_selector" type="select" label="Do you want to tweak effort options?" help="See &quot;Effort Options&quot; section of Help below for information">
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391 <option value="yes">Yes</option>
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392 <option value="no" selected="true">No</option>
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393 </param>
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394 <when value="yes">
3
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395 <param name="D" type="integer" value="15" min="0" label="Attempt that many consecutive seed extension attempts to `fail` before Bowtie 2 moves on, using the alignments found so far" help="-D; A seed extension `fails` if it does not yield a new best or a new second-best alignment. This limit is automatically adjusted up when -k or -a are specified. Default=15"/>
2
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396 <param name="R" type="integer" value="2" min="0" label="Set the maximum number of times Bowtie 2 will `re-seed` reads with repetitive seeds" help="When `re-seeding`, Bowtie 2 simply chooses a new set of reads (same length, same number of mismatches allowed) at different offsets and searches for more alignments. A read is considered to have repetitive seeds if the total number of seed hits divided by the number of seeds that aligned at least once is greater than 300. Default=2"/>
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397 </when>
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398 <when value="no">
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parents: 0
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399 <!-- do nothing -->
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400 </when>
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401 </conditional>
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402
2
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403 <conditional name="sam_options">
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404 <param name="sam_options_selector" type="select" label="Do you want to tweak SAM/BAM Options?" help="See &quot;Output Options&quot; section of Help below for information">
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405 <option value="yes">Yes</option>
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406 <option value="no" selected="true">No</option>
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407 </param>
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408 <when value="yes">
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409 <param name="no_unal" type="boolean" truevalue="--no-unal" falsevalue="" label="Suppress SAM records for reads that failed to align" help="--no-unal; Default=False"/>
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410 <param name="omit_sec_seq" type="boolean" truevalue="--omit-sec-seq" falsevalue="" label="Suppress SEQ and QUAL strings for secondary alignments" help="--omit-sec-seq; Default=False"/>
2
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411 </when>
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412 <when value="no">
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413 <!-- do nothing -->
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414 </when>
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415 </conditional>
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416 <conditional name="other_options">
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417 <param name="other_options_selector" type="select" label="Do you want to tweak Other Options?" help="See &quot;Other Options&quot; section of Help below for information">
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418 <option value="yes">Yes</option>
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419 <option value="no" selected="true">No</option>
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420 </param>
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421 <when value="yes">
7
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422 <param name="reorder" type="boolean" truevalue="--reorder" falsevalue="" label="Guarantee that output SAM records are printed in an order corresponding to the order of the reads in the original input file" help="--reorder; Default=False"/>
2
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423 <param name="seed" type="integer" value="0" min="0" label="Use this number as the seed for pseudo-random number generator" help="--seed; Default=0"/>
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424 <param name="non_deterministic" type="boolean" truevalue="--non-deterministic" falsevalue="" label="Re-initialize the pseudo-random generator for each read using the current time" help="--non-deterministic; see Help below for explanation of this option; default=False"/>
2
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425 </when>
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426 <when value="no">
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parents: 0
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427 <!-- do nothing -->
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428 </when>
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429 </conditional>
7
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430 <param name="sam_opt" type="boolean" truevalue="true" falsevalue="false" label="Would you like the output to be a SAM file" help="By default, the output from this Bowtie2 wrapper is a sorted BAM file."/>
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431 </when>
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432 </conditional>
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433 </inputs>
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434
96d2e31a3938 Imported from capsule None
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435 <!-- define outputs -->
2
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436
0
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437 <outputs>
2
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438
0
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439 <data format="fastqsanger" name="output_unaligned_reads_l" label="${tool.name} on ${on_string}: unaligned reads (L)" >
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440 <filter>library['unaligned_file'] is True</filter>
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96d2e31a3938 Imported from capsule None
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441 <actions>
7
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442 <conditional name="library.type">
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443 <when value="single">
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444 <action type="format">
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445 <option type="from_param" name="library.input_1" param_attribute="ext" />
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446 </action>
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447 </when>
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448 <when value="paired">
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449 <action type="format">
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450 <option type="from_param" name="library.input_1" param_attribute="ext" />
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451 </action>
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452 </when>
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453 <when value="paired_collection">
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454 <action type="format">
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455 <option type="from_param" name="library.input_1" param_attribute="forward.ext" />
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456 </action>
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457 </when>
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458 </conditional>
0
96d2e31a3938 Imported from capsule None
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459 </actions>
96d2e31a3938 Imported from capsule None
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460 </data>
96d2e31a3938 Imported from capsule None
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461 <data format="fastqsanger" name="output_unaligned_reads_r" label="${tool.name} on ${on_string}: unaligned reads (R)">
2
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462 <filter>( library['type'] == "paired" or library['type'] == "paired_collection" ) and library['unaligned_file'] is True</filter>
0
96d2e31a3938 Imported from capsule None
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parents:
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463 <actions>
7
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464 <conditional name="library.type">
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465 <when value="paired">
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466 <action type="format">
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467 <option type="from_param" name="library.input_2" param_attribute="ext" />
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468 </action>
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469 </when>
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470 <when value="paired_collection">
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471 <action type="format">
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472 <option type="from_param" name="library.input_1" param_attribute="reverse.ext" />
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473 </action>
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474 </when>
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475 </conditional>
0
96d2e31a3938 Imported from capsule None
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476 </actions>
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477 </data>
2
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478
3
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479 <data format="bam" name="output" label="${tool.name} on ${on_string}: aligned reads (sorted BAM)">
7
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480 <filter>analysis_type['analysis_type_selector'] == "simple" or analysis_type['sam_opt'] is False</filter>
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481 <actions>
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482 <conditional name="reference_genome.source">
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483 <when value="indexed">
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484 <action type="metadata" name="dbkey">
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485 <option type="from_data_table" name="bowtie2_indexes" column="1" offset="0">
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486 <filter type="param_value" column="0" value="#" compare="startswith" keep="False"/>
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487 <filter type="param_value" ref="reference_genome.index" column="0"/>
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488 </option>
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489 </action>
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490 </when>
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491 <when value="history">
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492 <action type="metadata" name="dbkey">
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493 <option type="from_param" name="reference_genome.own_file" param_attribute="dbkey" />
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494 </action>
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495 </when>
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496 </conditional>
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497 </actions>
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498 </data>
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499
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500 <data format="sam" name="output_sam" label="${tool.name} on ${on_string}: aligned reads (SAM)">
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501 <filter>analysis_type['analysis_type_selector'] == "full" and analysis_type['sam_opt'] is True</filter>
0
96d2e31a3938 Imported from capsule None
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502 <actions>
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503 <conditional name="reference_genome.source">
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504 <when value="indexed">
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505 <action type="metadata" name="dbkey">
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506 <option type="from_data_table" name="bowtie2_indexes" column="1" offset="0">
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507 <filter type="param_value" column="0" value="#" compare="startswith" keep="False"/>
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508 <filter type="param_value" ref="reference_genome.index" column="0"/>
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509 </option>
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510 </action>
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511 </when>
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512 <when value="history">
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513 <action type="metadata" name="dbkey">
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514 <option type="from_param" name="reference_genome.own_file" param_attribute="dbkey" />
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515 </action>
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516 </when>
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517 </conditional>
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518 </actions>
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519 </data>
3
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520
0
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521 </outputs>
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522
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523 <tests>
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524 <test>
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525 <!-- basic test on single paired default run -->
2
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526 <param name="type" value="paired"/>
0
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527 <param name="selection" value="no"/>
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528 <param name="paired_options_selector" value="no"/>
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529 <param name="unaligned_file" value="false"/>
2
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530 <param name="analysis_type_selector" value="simple"/>
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531 <param name="source" value="history" />
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532 <param name="input_1" value="bowtie2-fq1.fq" ftype="fastqsanger"/>
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533 <param name="input_2" value="bowtie2-fq2.fq" ftype="fastqsanger"/>
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534 <param name="own_file" value="bowtie2-ref.fasta" />
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535 <output name="output" file="bowtie2-test1.bam" ftype="bam" lines_diff="2"/>
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536 </test>
5
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537 <test>
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538 <!-- basic test on single paired default run -->
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539 <param name="type" value="paired"/>
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540 <param name="selection" value="no"/>
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541 <param name="paired_options_selector" value="no"/>
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542 <param name="unaligned_file" value="false"/>
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543 <param name="analysis_type_selector" value="simple"/>
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544 <param name="rg_selector" value="set"/>
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545 <param name="ID" value="rg1"/>
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546 <param name="PL" value="CAPILLARY"/>
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547 <param name="source" value="history" />
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548 <param name="input_1" value="bowtie2-fq1.fq" ftype="fastqsanger"/>
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549 <param name="input_2" value="bowtie2-fq2.fq" ftype="fastqsanger"/>
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550 <param name="own_file" value="bowtie2-ref.fasta" />
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551 <output name="output" file="bowtie2-test2.bam" ftype="bam" lines_diff="2"/>
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552 </test>
0
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553 </tests>
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554
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555 <help>
2
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556
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557 **Bowtie2 Overview**
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558
2
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559 Bowtie2_ is an ultrafast and memory-efficient tool for aligning sequencing reads to long reference sequences. It is particularly good at aligning reads of about 50 up to 100s or 1,000s of characters to relatively long (e.g. mammalian) genomes. Bowtie 2 supports gapped, local, and paired-end alignment modes. Galaxy wrapper for Bowtie 2 outputs alignments in `BAM format`_, enabling interoperation with a large number of other tools available at this site.
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560 Majority of information in this page is derived from an excellent `Bowtie2 manual`_ written by Ben Langmead.
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561
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562 .. _Bowtie2: http://bowtie-bio.sourceforge.net/bowtie2/
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563 .. _`Bowtie2 manual`: http://bowtie-bio.sourceforge.net/bowtie2/manual.shtml
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564 .. _`BAM format`: http://samtools.github.io/hts-specs/SAMv1.pdf
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565
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566 -----
0
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567
2
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568 **Selecting reference genomes for Bowtie2**
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569
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570 Galaxy wrapper for Bowtie2 allows you select between precomputed and user-defined indices for reference genomes using **Will you select a reference genome from your history or use a built-in index?** flag. This flag has two options:
0
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571
2
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572 1. **Use a built-in genome index** - when selected (this is default), Galaxy provides the user with **Select reference genome index** dropdown. Genomes listed in this dropdown have been pre-indexed with bowtie2-build utility and are ready to be mapped against.
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573 2. **Use a genome from the history and build index** - when selected, Galaxy provides the user with **Select reference genome sequence** dropdown. This dropdown is populated by all FASTA formatted files listed in your current history. If your genome of interest is uploaded into history it will be shown there. Selecting a genome from this dropdown will cause Galaxy to first transparently index it using bowtie2-build command, and then run mapping with bowtie2.
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574
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575 If your genome of interest is not listed here you have two choices:
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576
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577 1. Contact galaxy team using **Help->Support** link at the top of the interface and let us know that an index needs to be added
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578 2. Upload your genome of interest as a FASTA file to Galaxy history and selected **Use a genome from the history and build index** option.
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579
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580 ------
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581
2
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582 .. class:: infomark
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583
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584 **Bowtie2 options**
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585
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586 Galaxy wrapper for Bowtie2 implements most but not all options available through the command line. Supported options are described below.
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587
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588 -----
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589
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590 **Inputs**
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591
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592 Bowtie 2 accepts files in Sanger FASTQ format (single or pair-end). Use the FASTQ Groomer to prepare your files.
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593
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594 ------
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595
2
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596 **Input options**::
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597
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598 -s/--skip &lt;int&gt;
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599 Skip (i.e. do not align) the first `&lt;int&gt;` reads or pairs in the input.
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600
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601 -u/--qupto &lt;int&gt;
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602 Align the first `&lt;int&gt;` reads or read pairs from the input (after the
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603 `-s`/`--skip` reads or pairs have been skipped), then stop. Default: no limit.
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604
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605 -5/--trim5 &lt;int&gt;
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606 Trim `&lt;int&gt;` bases from 5' (left) end of each read before alignment (default: 0).
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607
2
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608 -3/--trim3 &lt;int&gt;
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609 Trim `&lt;int&gt;` bases from 3' (right) end of each read before alignment (default: 0).
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610
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611 --phred33
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612 Input qualities are ASCII chars equal to the Phred quality plus 33. This is
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613 also called the "Phred+33" encoding, which is used by the very latest Illumina
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614 pipelines.
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615
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616 --phred64
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617 Input qualities are ASCII chars equal to the Phred quality plus 64. This is
2
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618 also called the "Phred+64" encoding.
0
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619
2
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620 --solexa-quals
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621 Convert input qualities from Solexa Phred quality (which can be negative) to
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622 Phred Phred quality (which can't). This scheme was used in older Illumina GA
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623 Pipeline versions (prior to 1.3). Default: off.
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624
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625 --int-quals
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626 Quality values are represented in the read input file as space-separated ASCII integers, e.g., `40 40 30 40`..., rather than ASCII characters, e.g., `II?I`....
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627 Integers are treated as being on the Phred quality scale unless
2
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628 `--solexa-quals` is also specified. Default: off.
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629
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630 ------
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631
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632 **Presets in `--end-to-end` mode**::
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633
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634 --very-fast
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635 Same as: `-D 5 -R 1 -N 0 -L 22 -i S,0,2.50`
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636
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637 --fast
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638 Same as: `-D 10 -R 2 -N 0 -L 22 -i S,0,2.50`
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639
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640 --sensitive
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641 Same as: `-D 15 -R 2 -L 22 -i S,1,1.15` (default in `--end-to-end` mode)
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642
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643 --very-sensitive
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644 Same as: `-D 20 -R 3 -N 0 -L 20 -i S,1,0.50`
0
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645
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646 ------
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647
2
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648 **Presets options in `--local` mode**::
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649
2
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650 --very-fast-local
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651 Same as: `-D 5 -R 1 -N 0 -L 25 -i S,1,2.00`
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652
2
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653 --fast-local
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654 Same as: `-D 10 -R 2 -N 0 -L 22 -i S,1,1.75`
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655
2
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656 --sensitive-local
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657 Same as: `-D 15 -R 2 -N 0 -L 20 -i S,1,0.75` (default in `--local` mode)
0
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658
2
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659 --very-sensitive-local
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660 Same as: `-D 20 -R 3 -N 0 -L 20 -i S,1,0.50`
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661
0
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662 ------
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663
2
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664 **Alignment options**::
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665
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666 -N &lt;int&gt;
3
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667 Sets the number of mismatches to allowed in a seed alignment during multiseed
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668 alignment. Can be set to 0 or 1. Setting this higher makes alignment slower
2
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669 (often much slower) but increases sensitivity. Default: 0.
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670
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671 -L &lt;int&gt;
3
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672 Sets the length of the seed substrings to align during multiseed alignment.
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673 Smaller values make alignment slower but more sensitive. Default: the
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674 `--sensitive` preset is used by default, which sets `-L` to 22 in
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675 `--end-to-end` mode and to 20 in `--local` mode.
2
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676
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677 -i &lt;func&gt;
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678 Sets a function governing the interval between seed substrings to use during
3
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diff changeset
679 multiseed alignment. For instance, if the read has 30 characers, and seed
2
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parents: 0
diff changeset
680 length is 10, and the seed interval is 6, the seeds extracted will be:
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parents: 0
diff changeset
681
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diff changeset
682 Read: TAGCTACGCTCTACGCTATCATGCATAAAC
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parents: 0
diff changeset
683 Seed 1 fw: TAGCTACGCT
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parents: 0
diff changeset
684 Seed 1 rc: AGCGTAGCTA
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diff changeset
685 Seed 2 fw: CGCTCTACGC
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parents: 0
diff changeset
686 Seed 2 rc: GCGTAGAGCG
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parents: 0
diff changeset
687 Seed 3 fw: ACGCTATCAT
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parents: 0
diff changeset
688 Seed 3 rc: ATGATAGCGT
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parents: 0
diff changeset
689 Seed 4 fw: TCATGCATAA
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parents: 0
diff changeset
690 Seed 4 rc: TTATGCATGA
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parents: 0
diff changeset
691
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parents: 0
diff changeset
692 Since it's best to use longer intervals for longer reads, this parameter sets
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parents: 0
diff changeset
693 the interval as a function of the read length, rather than a single
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parents: 0
diff changeset
694 one-size-fits-all number. For instance, specifying `-i S,1,2.5` sets the
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parents: 0
diff changeset
695 interval function `f` to `f(x) = 1 + 2.5 * sqrt(x)`, where x is the read length.
3
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parents: 2
diff changeset
696 If the function returns a result less than
2
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parents: 0
diff changeset
697 1, it is rounded up to 1. Default: the `--sensitive` preset is used by
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parents: 0
diff changeset
698 default, which sets `-i` to `S,1,1.15` in `--end-to-end` mode to `-i S,1,0.75`
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parents: 0
diff changeset
699 in `--local` mode.
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parents: 0
diff changeset
700
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parents: 0
diff changeset
701 --n-ceil &lt;func&gt;
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parents: 0
diff changeset
702 Sets a function governing the maximum number of ambiguous characters (usually
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parents: 0
diff changeset
703 `N`s and/or `.`s) allowed in a read as a function of read length. For instance,
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devteam
parents: 0
diff changeset
704 specifying `-L,0,0.15` sets the N-ceiling function `f` to `f(x) = 0 + 0.15 * x`,
3
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devteam
parents: 2
diff changeset
705 where x is the read length. Reads exceeding this ceiling are filtered out.
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parents: 2
diff changeset
706 Default: `L,0,0.15`.
2
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parents: 0
diff changeset
707
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parents: 0
diff changeset
708 --dpad &lt;int&gt;
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parents: 0
diff changeset
709 "Pads" dynamic programming problems by `&lt;int&gt;` columns on either side to allow
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parents: 0
diff changeset
710 gaps. Default: 15.
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parents: 0
diff changeset
711
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parents: 0
diff changeset
712 --gbar &lt;int&gt;
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parents: 0
diff changeset
713 Disallow gaps within `&lt;int&gt;` positions of the beginning or end of the read.
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parents: 0
diff changeset
714 Default: 4.
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parents: 0
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715
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parents: 0
diff changeset
716 --ignore-quals
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parents: 0
diff changeset
717 When calculating a mismatch penalty, always consider the quality value at the
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parents: 0
diff changeset
718 mismatched position to be the highest possible, regardless of the actual value.
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parents: 0
diff changeset
719 I.e. input is treated as though all quality values are high. This is also the
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parents: 0
diff changeset
720 default behavior when the input doesn't specify quality values (e.g. in `-f`,
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parents: 0
diff changeset
721 `-r`, or `-c` modes).
0
96d2e31a3938 Imported from capsule None
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722
2
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parents: 0
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723 --nofw/--norc
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parents: 0
diff changeset
724 If `--nofw` is specified, `bowtie2` will not attempt to align unpaired reads to
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parents: 0
diff changeset
725 the forward (Watson) reference strand. If `--norc` is specified, `bowtie2` will
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parents: 0
diff changeset
726 not attempt to align unpaired reads against the reverse-complement (Crick)
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parents: 0
diff changeset
727 reference strand. In paired-end mode, `--nofw` and `--norc` pertain to the
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diff changeset
728 fragments; i.e. specifying `--nofw` causes `bowtie2` to explore only those
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parents: 0
diff changeset
729 paired-end configurations corresponding to fragments from the reverse-complement
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parents: 0
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730 (Crick) strand. Default: both strands enabled.
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parents: 0
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731
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parents: 0
diff changeset
732 --no-1mm-upfront
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parents: 0
diff changeset
733 By default, Bowtie 2 will attempt to find either an exact or a 1-mismatch
3
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parents: 2
diff changeset
734 end-to-end alignment for the read *before* trying the multiseed heuristic. Such
2
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parents: 0
diff changeset
735 alignments can be found very quickly, and many short read alignments have exact or
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parents: 0
diff changeset
736 near-exact end-to-end alignments. However, this can lead to unexpected
3
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parents: 2
diff changeset
737 alignments when the user also sets options governing the multiseed heuristic,
2
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parents: 0
diff changeset
738 like `-L` and `-N`. For instance, if the user specifies `-N 0` and `-L` equal
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parents: 0
diff changeset
739 to the length of the read, the user will be surprised to find 1-mismatch alignments
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parents: 0
diff changeset
740 reported. This option prevents Bowtie 2 from searching for 1-mismatch end-to-end
3
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parents: 2
diff changeset
741 alignments before using the multiseed heuristic, which leads to the expected
2
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parents: 0
diff changeset
742 behavior when combined with options such as `-L` and `-N`. This comes at the
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parents: 0
diff changeset
743 expense of speed.
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parents: 0
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744
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parents: 0
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745 --end-to-end
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parents: 0
diff changeset
746 In this mode, Bowtie 2 requires that the entire read align from one end to the
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parents: 0
diff changeset
747 other, without any trimming (or "soft clipping") of characters from either end.
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parents: 0
diff changeset
748 The match bonus `--ma` always equals 0 in this mode, so all alignment scores
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parents: 0
diff changeset
749 are less than or equal to 0, and the greatest possible alignment score is 0.
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parents: 0
diff changeset
750 This is mutually exclusive with `--local`. `--end-to-end` is the default mode.
0
96d2e31a3938 Imported from capsule None
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parents:
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751
2
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parents: 0
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752 --local
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parents: 0
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753 In this mode, Bowtie 2 does not require that the entire read align from one end
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parents: 0
diff changeset
754 to the other. Rather, some characters may be omitted ("soft clipped") from the
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parents: 0
diff changeset
755 ends in order to achieve the greatest possible alignment score. The match bonus
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parents: 0
diff changeset
756 `--ma` is used in this mode, and the best possible alignment score is equal to
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parents: 0
diff changeset
757 the match bonus (`--ma`) times the length of the read. Specifying `--local`
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parents: 0
diff changeset
758 and one of the presets (e.g. `--local --very-fast`) is equivalent to specifying
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parents: 0
diff changeset
759 the local version of the preset (`--very-fast-local`). This is mutually
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parents: 0
diff changeset
760 exclusive with `--end-to-end`. `--end-to-end` is the default mode.
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parents: 0
diff changeset
761
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parents: 0
diff changeset
762 -----
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parents: 0
diff changeset
763
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parents: 0
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764 **Scoring options**::
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parents: 0
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765
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parents: 0
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766 --ma &lt;int&gt;
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parents: 0
diff changeset
767 Sets the match bonus. In `--local` mode `&lt;int&gt;` is added to the alignment
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parents: 0
diff changeset
768 score for each position where a read character aligns to a reference character
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parents: 0
diff changeset
769 and the characters match. Not used in `--end-to-end` mode. Default: 2.
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parents: 0
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770
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parents: 0
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771 --mp MX,MN
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diff changeset
772 Sets the maximum (`MX`) and minimum (`MN`) mismatch penalties, both integers. A
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parents: 0
diff changeset
773 number less than or equal to `MX` and greater than or equal to `MN` is
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parents: 0
diff changeset
774 subtracted from the alignment score for each position where a read character
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parents: 0
diff changeset
775 aligns to a reference character, the characters do not match, and neither is an
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parents: 0
diff changeset
776 `N`. If `--ignore-quals` is specified, the number subtracted quals `MX`.
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parents: 0
diff changeset
777 Otherwise, the number subtracted is `MN + floor( (MX-MN)(MIN(Q, 40.0)/40.0) )`
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parents: 0
diff changeset
778 where Q is the Phred quality value. Default: `MX` = 6, `MN` = 2.
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parents: 0
diff changeset
779
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parents: 0
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780 --np &lt;int&gt;
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parents: 0
diff changeset
781 Sets penalty for positions where the read, reference, or both, contain an
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parents: 0
diff changeset
782 ambiguous character such as `N`. Default: 1.
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parents: 0
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783
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parents: 0
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784 --rdg &lt;int1&gt;,&lt;int2&gt;
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parents: 0
diff changeset
785 Sets the read gap open (`&lt;int1&gt;`) and extend (`&lt;int2&gt;`) penalties. A read gap of
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parents: 0
diff changeset
786 length N gets a penalty of `&lt;int1&gt;` + N * `&lt;int2&gt;`. Default: 5, 3.
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parents: 0
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787
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diff changeset
788 --rfg &lt;int1&gt;,&lt;int2&gt;
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parents: 0
diff changeset
789 Sets the reference gap open (`&lt;int1&gt;`) and extend (`&lt;int2&gt;`) penalties. A
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parents: 0
diff changeset
790 reference gap of length N gets a penalty of `&lt;int1&gt;` + N * `&lt;int2&gt;`. Default:
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parents: 0
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791 5, 3.
0
96d2e31a3938 Imported from capsule None
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792
2
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parents: 0
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793 --score-min &lt;func&gt;
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parents: 0
diff changeset
794 Sets a function governing the minimum alignment score needed for an alignment to
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parents: 0
diff changeset
795 be considered "valid" (i.e. good enough to report). This is a function of read
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parents: 0
diff changeset
796 length. For instance, specifying `L,0,-0.6` sets the minimum-score function `f`
3
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parents: 2
diff changeset
797 to `f(x) = 0 + -0.6 * x`, where `x` is the read length. The default in `--end-to-end` mode is `L,-0.6,-0.6` and
2
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diff changeset
798 the default in `--local` mode is `G,20,8`.
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parents: 0
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799
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parents: 0
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800 -----
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parents: 0
diff changeset
801
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parents: 0
diff changeset
802 **Reporting options**::
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parents: 0
diff changeset
803
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parents: 0
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804 -k &lt;int&gt;
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parents: 0
diff changeset
805 By default, `bowtie2` searches for distinct, valid alignments for each read.
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parents: 0
diff changeset
806 When it finds a valid alignment, it continues looking for alignments that are
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parents: 0
diff changeset
807 nearly as good or better. The best alignment found is reported (randomly
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parents: 0
diff changeset
808 selected from among best if tied). Information about the best alignments is
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parents: 0
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809 used to estimate mapping quality and to set SAM optional fields, such as
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parents: 0
diff changeset
810 `AS:i` and `XS:i`.
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parents: 0
diff changeset
811
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parents: 0
diff changeset
812 When `-k` is specified, however, `bowtie2` behaves differently. Instead, it
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parents: 0
diff changeset
813 searches for at most `&lt;int&gt;` distinct, valid alignments for each read. The
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parents: 0
diff changeset
814 search terminates when it can't find more distinct valid alignments, or when it
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parents: 0
diff changeset
815 finds `&lt;int&gt;`, whichever happens first. All alignments found are reported in
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parents: 0
diff changeset
816 descending order by alignment score. The alignment score for a paired-end
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parents: 0
diff changeset
817 alignment equals the sum of the alignment scores of the individual mates. Each
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parents: 0
diff changeset
818 reported read or pair alignment beyond the first has the SAM 'secondary' bit
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parents: 0
diff changeset
819 (which equals 256) set in its FLAGS field. For reads that have more than
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parents: 0
diff changeset
820 `&lt;int&gt;` distinct, valid alignments, `bowtie2` does not guarantee that the
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parents: 0
diff changeset
821 `&lt;int&gt;` alignments reported are the best possible in terms of alignment score.
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parents: 0
diff changeset
822 `-k` is mutually exclusive with `-a`.
0
96d2e31a3938 Imported from capsule None
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parents:
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823
2
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parents: 0
diff changeset
824 Note: Bowtie 2 is not designed with large values for `-k` in mind, and when
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parents: 0
diff changeset
825 aligning reads to long, repetitive genomes large `-k` can be very, very slow.
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parents: 0
diff changeset
826
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parents: 0
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827 -a
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parents: 0
diff changeset
828 Like `-k` but with no upper limit on number of alignments to search for. `-a`
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parents: 0
diff changeset
829 is mutually exclusive with `-k`.
c1ec08cb34f9 Uploaded
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parents: 0
diff changeset
830
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parents: 0
diff changeset
831 Note: Bowtie 2 is not designed with `-a` mode in mind, and when
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parents: 0
diff changeset
832 aligning reads to long, repetitive genomes this mode can be very, very slow.
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parents: 0
diff changeset
833
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parents: 0
diff changeset
834 -----
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parents: 0
diff changeset
835
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parents: 0
diff changeset
836 **Effort options**::
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parents: 0
diff changeset
837
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parents: 0
diff changeset
838 -D &lt;int&gt;
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parents: 0
diff changeset
839 Up to `&lt;int&gt;` consecutive seed extension attempts can "fail" before Bowtie 2
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parents: 0
diff changeset
840 moves on, using the alignments found so far. A seed extension "fails" if it
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parents: 0
diff changeset
841 does not yield a new best or a new second-best alignment. This limit is
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parents: 0
diff changeset
842 automatically adjusted up when -k or -a are specified. Default: 15.
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parents: 0
diff changeset
843
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parents: 0
diff changeset
844 -R &lt;int&gt;
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parents: 0
diff changeset
845 `&lt;int&gt;` is the maximum number of times Bowtie 2 will "re-seed" reads with
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parents: 0
diff changeset
846 repetitive seeds. When "re-seeding," Bowtie 2 simply chooses a new set of reads
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parents: 0
diff changeset
847 (same length, same number of mismatches allowed) at different offsets and
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parents: 0
diff changeset
848 searches for more alignments. A read is considered to have repetitive seeds if
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parents: 0
diff changeset
849 the total number of seed hits divided by the number of seeds that aligned at
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parents: 0
diff changeset
850 least once is greater than 300. Default: 2.
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parents: 0
diff changeset
851
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parents: 0
diff changeset
852 -----
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parents: 0
diff changeset
853
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parents: 0
diff changeset
854 **Paired-end options**::
0
96d2e31a3938 Imported from capsule None
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parents:
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855
2
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parents: 0
diff changeset
856 -I/--minins &lt;int&gt;
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parents: 0
diff changeset
857 The minimum fragment length for valid paired-end alignments. E.g. if `-I 60` is
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parents: 0
diff changeset
858 specified and a paired-end alignment consists of two 20-bp alignments in the
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parents: 0
diff changeset
859 appropriate orientation with a 20-bp gap between them, that alignment is
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parents: 0
diff changeset
860 considered valid (as long as `-X` is also satisfied). A 19-bp gap would not
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parents: 0
diff changeset
861 be valid in that case. If trimming options `-3` or `-5` are also used, the
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parents: 0
diff changeset
862 `-I` constraint is applied with respect to the untrimmed mates.
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parents: 0
diff changeset
863
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parents: 0
diff changeset
864 The larger the difference between `-I` and `-X`, the slower Bowtie 2 will
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865 run. This is because larger differences bewteen `-I` and `-X` require that
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diff changeset
866 Bowtie 2 scan a larger window to determine if a concordant alignment exists.
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diff changeset
867 For typical fragment length ranges (200 to 400 nucleotides), Bowtie 2 is very
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868 efficient.
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869
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870 Default: 0 (essentially imposing no minimum)
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diff changeset
871
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parents: 0
diff changeset
872 -X/--maxins &lt;int&gt;
c1ec08cb34f9 Uploaded
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diff changeset
873 The maximum fragment length for valid paired-end alignments. E.g. if `-X 100`
c1ec08cb34f9 Uploaded
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diff changeset
874 is specified and a paired-end alignment consists of two 20-bp alignments in the
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875 proper orientation with a 60-bp gap between them, that alignment is considered
c1ec08cb34f9 Uploaded
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876 valid (as long as `-I` is also satisfied). A 61-bp gap would not be valid in
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diff changeset
877 that case. If trimming options `-3` or `-5` are also used, the `-X`
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878 constraint is applied with respect to the untrimmed mates, not the trimmed
c1ec08cb34f9 Uploaded
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879 mates.
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diff changeset
880
c1ec08cb34f9 Uploaded
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diff changeset
881 The larger the difference between `-I` and `-X`, the slower Bowtie 2 will
c1ec08cb34f9 Uploaded
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diff changeset
882 run. This is because larger differences bewteen `-I` and `-X` require that
c1ec08cb34f9 Uploaded
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parents: 0
diff changeset
883 Bowtie 2 scan a larger window to determine if a concordant alignment exists.
c1ec08cb34f9 Uploaded
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parents: 0
diff changeset
884 For typical fragment length ranges (200 to 400 nucleotides), Bowtie 2 is very
c1ec08cb34f9 Uploaded
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parents: 0
diff changeset
885 efficient.
c1ec08cb34f9 Uploaded
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diff changeset
886
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887 Default: 500.
0
96d2e31a3938 Imported from capsule None
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888
2
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889 --fr/--rf/--ff
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diff changeset
890 The upstream/downstream mate orientations for a valid paired-end alignment
c1ec08cb34f9 Uploaded
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diff changeset
891 against the forward reference strand. E.g., if `--fr` is specified and there is
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diff changeset
892 a candidate paired-end alignment where mate 1 appears upstream of the reverse
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diff changeset
893 complement of mate 2 and the fragment length constraints (`-I` and `-X`) are
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894 met, that alignment is valid. Also, if mate 2 appears upstream of the reverse
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diff changeset
895 complement of mate 1 and all other constraints are met, that too is valid.
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896 `--rf` likewise requires that an upstream mate1 be reverse-complemented and a
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diff changeset
897 downstream mate2 be forward-oriented. ` --ff` requires both an upstream mate 1
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diff changeset
898 and a downstream mate 2 to be forward-oriented. Default: `--fr` (appropriate
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parents: 0
diff changeset
899 for Illumina's Paired-end Sequencing Assay).
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diff changeset
900
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901 --no-mixed
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diff changeset
902 By default, when `bowtie2` cannot find a concordant or discordant alignment for
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903 a pair, it then tries to find alignments for the individual mates. This option
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904 disables that behavior.
0
96d2e31a3938 Imported from capsule None
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parents:
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905
2
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906 --no-discordant
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907 By default, `bowtie2` looks for discordant alignments if it cannot find any
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908 concordant alignments. A discordant alignment is an alignment where both mates
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diff changeset
909 align uniquely, but that does not satisfy the paired-end constraints
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910 (`--fr`/`--rf`/`--ff`, `-I`, `-X`). This option disables that behavior.
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diff changeset
911
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912 --dovetail
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diff changeset
913 If the mates "dovetail", that is if one mate alignment extends past the
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parents: 0
diff changeset
914 beginning of the other such that the wrong mate begins upstream, consider that
3
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diff changeset
915 to be concordant. Default: mates cannot dovetail in a concordant alignment.
2
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diff changeset
916
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917 --no-contain
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diff changeset
918 If one mate alignment contains the other, consider that to be non-concordant.
3
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diff changeset
919 Default: a mate can contain the other in a concordant alignment.
2
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parents: 0
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920
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921 --no-overlap
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diff changeset
922 If one mate alignment overlaps the other at all, consider that to be
3
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parents: 2
diff changeset
923 non-concordant. Default: mates can overlap in a concordant alignment.
2
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diff changeset
924
0
96d2e31a3938 Imported from capsule None
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925 ------
96d2e31a3938 Imported from capsule None
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parents:
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926
2
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diff changeset
927 **SAM options**::
0
96d2e31a3938 Imported from capsule None
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parents:
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928
2
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diff changeset
929 --rg-id &lt;text&gt;
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diff changeset
930 Set the read group ID to `&lt;text&gt;`. This causes the SAM `@RG` header line to be
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diff changeset
931 printed, with `&lt;text&gt;` as the value associated with the `ID:` tag. It also
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diff changeset
932 causes the `RG:Z:` extra field to be attached to each SAM output record, with
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933 value set to `&lt;text&gt;`.
0
96d2e31a3938 Imported from capsule None
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parents:
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934
2
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935 --rg &lt;text&gt;
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diff changeset
936 Add `&lt;text&gt;` (usually of the form `TAG:VAL`, e.g. `SM:Pool1`) as a field on the
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937 `@RG` header line. Note: in order for the `@RG` line to appear, `--rg-id`
3
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parents: 2
diff changeset
938 must also be specified. This is because the `ID` tag is required by the SAM
ceb6467e276c Uploaded
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parents: 2
diff changeset
939 Specification. Specify `--rg` multiple times to set multiple fields. See the
ceb6467e276c Uploaded
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parents: 2
diff changeset
940 SAM Specification for details about what fields are legal.
0
96d2e31a3938 Imported from capsule None
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parents:
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941
2
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942 --omit-sec-seq
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parents: 0
diff changeset
943 When printing secondary alignments, Bowtie 2 by default will write out the `SEQ`
c1ec08cb34f9 Uploaded
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944 and `QUAL` strings. Specifying this option causes Bowtie 2 to print an asterix
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945 in those fields instead.
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946
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parents: 0
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947 -----
0
96d2e31a3938 Imported from capsule None
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948
2
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949 **Other options**::
0
96d2e31a3938 Imported from capsule None
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950
2
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951 --reorder
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diff changeset
952 Guarantees that output SAM records are printed in an order corresponding to the
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parents: 0
diff changeset
953 order of the reads in the original input file, even when `-p` is set greater
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954 than 1. Specifying `--reorder` and setting `-p` greater than 1 causes Bowtie
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955 2 to run somewhat slower and use somewhat more memory then if `--reorder` were
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956 not specified. Has no effect if `-p` is set to 1, since output order will
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957 naturally correspond to input order in that case.
0
96d2e31a3938 Imported from capsule None
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958
2
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959 --seed &lt;int&gt;
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960 Use `&lt;int&gt;` as the seed for pseudo-random number generator. Default: 0.
0
96d2e31a3938 Imported from capsule None
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parents:
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961
2
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962 --non-deterministic
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parents: 0
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963 Normally, Bowtie 2 re-initializes its pseudo-random generator for each read. It
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parents: 0
diff changeset
964 seeds the generator with a number derived from (a) the read name, (b) the
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parents: 0
diff changeset
965 nucleotide sequence, (c) the quality sequence, (d) the value of the `--seed`
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diff changeset
966 option. This means that if two reads are identical (same name, same
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967 nucleotides, same qualities) Bowtie 2 will find and report the same alignment(s)
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968 for both, even if there was ambiguity. When `--non-deterministic` is specified,
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969 Bowtie 2 re-initializes its pseudo-random generator for each read using the
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970 current time. This means that Bowtie 2 will not necessarily report the same
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diff changeset
971 alignment for two identical reads. This is counter-intuitive for some users,
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972 but might be more appropriate in situations where the input consists of many
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973 identical reads.
0
96d2e31a3938 Imported from capsule None
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974
96d2e31a3938 Imported from capsule None
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975 </help>
2
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976 <citations>
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977 <citation type="doi">10.1186/gb-2009-10-3-r25</citation>
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parents: 0
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978 <citation type="doi">10.1038/nmeth.1923</citation>
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979 </citations>
0
96d2e31a3938 Imported from capsule None
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980 </tool>