Mercurial > repos > estrain > flye_cpu15mem59

annotate flye_cpu15mem59.xml @ 1:ab6ffc360b78 draft

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author estrain
date Fri, 13 Mar 2026 11:36:25 +0000
parents e1e6ef58f334
children 7049afca140c
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1 <tool id="flye" name="Flye" version="@TOOL_VERSION@+galaxy@SUFFIX_VERSION@" profile="20.01">
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2 <description>de novo assembler for single molecule sequencing reads</description>
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3 <macros>
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4 <import>macros.xml</import>
1
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5
0
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6 </macros>
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7 <expand macro="edam_ontology"/>
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8 <expand macro="xrefs"/>
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9 <expand macro="requirements" />
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10 <version_command>flye --version</version_command>
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11 <command detect_errors="exit_code"><![CDATA[
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12 #if $input.is_of_type('fastqsanger', 'fastq'):
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13 #set ext = 'fastq'
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14 #elif $input.is_of_type('fastqsanger.gz', 'fastq.gz'):
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15 #set ext = 'fastq.gz'
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16 #elif $input.is_of_type('fasta.gz'):
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17 #set ext = 'fasta.gz'
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18 #elif $input.is_of_type('fasta'):
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19 #set ext = 'fasta'
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20 #else:
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21 #set ext = 'dat'
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22 #end if
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23
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24 ln -sf '$input' ./input_0.${ext} &&
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25 flye $mode_conditional.mode ./input_0.${ext}
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26 -o out_dir
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27 -t \${GALAXY_SLOTS:-4}
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28 -i $iterations
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29 #if $mode_conditional.mode == '--pacbio-hifi' and $mode_conditional.hifi_error:
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30 --hifi-error $mode_conditional.hifi_error
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31 #end if
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32 #if $min_overlap:
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33 -m $min_overlap
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34 #end if
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35 #if $asm.asm_select == 'true':
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36 --asm-coverage $asm.asm_coverage
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37 -g '$asm.genome_size'
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38 #end if
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39 #if $meta:
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40 $meta
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41 #end if
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42 #if $scaffold:
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43 $scaffold
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44 #end if
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45 #if $no_alt_contigs:
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46 $no_alt_contigs
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47 #end if
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48 ]]></command>
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49 <inputs>
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50 <param name="input" type="data" format="fasta,fasta.gz,fastq,fastq.gz,fastqsanger.gz,fastqsanger" label="Read file" />
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51 <conditional name="mode_conditional">
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52 <param name="mode" type="select" label="Mode">
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53 <option value="--nano-raw">Nanopore raw (--nano-raw)</option>
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54 <option value="--nano-corr">Nanopore corrected (--nano-corr)</option>
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55 <option value="--nano-hq">Nanopore HQ (--nano-hq)</option>
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56 <option value="--pacbio-raw">PacBio raw (--pacbio-raw)</option>
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57 <option value="--pacbio-corr">PacBio corrected (--pacbio-corr)</option>
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58 <option value="--pacbio-hifi">PacBio HiFi (--pacbio-hifi)</option>
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59 </param>
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60 <when value="--nano-raw"/>
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61 <when value="--nano-corr"/>
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62 <when value="--nano-hq"/>
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63 <when value="--pacbio-raw"/>
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64 <when value="--pacbio-corr"/>
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65 <when value="--pacbio-hifi">
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66 <param argument="--hifi-error" type="float" min="0" max="1" optional="true" label="Expected HiFi reads error rate" help="Default: 0.01"/>
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67 </when>
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68 </conditional>
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69 <param argument="--iterations" type="integer" value="1" label="Number of polishing iterations"
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70 help="Polishing is performed as the final assembly stage. By default, Flye runs one polishing iteration. Additional iterations
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71 might correct a small number of extra errors (due to improvements on how reads may align to the corrected assembly). If the
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72 parameter is set to 0, the polishing is not performed"/>
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73 <param argument="--min-overlap" type="integer" min="1000" max="10000" optional="true" label="Minimum overlap between reads"
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74 help="This sets a minimum overlap length for two reads to be considered overlapping. By default it is chosen
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75 automatically based on the read length distribution (reads N90) and does not require manual setting. Typical
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76 value is 3k-5k (and down to 1k for datasets with shorter read length). Intuitively, we want to set this
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77 parameter as high as possible, so the repeat graph is less tangled. However, higher values might lead to assembly gaps.
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78 In some rare cases it makes sense to manually increase minimum overlap for assemblies of big genomes with long reads and high coverage." />
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79 <param argument="--keep-haplotypes" type="boolean" truevalue="--keep-haplotypes" falsevalue="" checked="False" label="Keep haplotypes"
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80 help="By default, Flye collapses graph structures caused by alternative haplotypes (bubbles, superbubbles, roundabouts) to produce longer
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81 consensus contigs. This option retains the alternative paths on the graph, producing less contigouos, but more detailed assembly."/>
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82 <param argument="--scaffold" type="boolean" truevalue="--scaffold" falsevalue="" label="Enable scaffolding using graph"
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83 help="Starting from the version 2.9 Flye does not perform scaffolding by default, which guarantees that all assembled sequences do not have any gaps" />
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84 <param argument="--meta" type="boolean" truevalue="--meta" falsevalue="" checked="False" label="Perform metagenomic assembly"
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85 help="It is designed for highly non-uniform coverage and is sensitive to underrepresented sequence at low coverage (as low as 2x).
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86 In some examples of simple metagenomes, we observed that the normal mode assembled more contigious bacterial
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87 consensus sequence, while the metagenome mode was slightly more fragmented, but revealed strain mixtures"/>
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88 <conditional name="asm">
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89 <param name="asm_select" type="select" label="Reduced contig assembly coverage" help="Typically, assemblies of large genomes at high coverage require a large amount of RAM. For high coverage assemblies, you can reduce memory usage by using only a subset of longest reads for initial contig extension stage (which is often the memory bottleneck)">
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90 <option value="true">Enable reduced coverage for initial disjointing assembly</option>
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91 <option value="false" selected="true">Disable reduced coverage for initial disjointing assembly</option>
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92 </param>
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93 <when value="true">
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94 <param argument="--asm-coverage" type="integer" min="0" value="30"
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95 label="Reduced coverage for initial disjointing assembly"
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96 help="This parameter specifies the target coverage of the longest reads. For a typical assembly, 30x is enough to produce good
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97 initial contigs. Regardless of this parameter, all reads will be used at the later pipeline stages."/>
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98 <param argument="--genome-size" type="text" optional="true" label="Estimated genome size"
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99 help="For example, 5m or 2.6g. No longer required as input. However, it must be used in conjunction with --asm-coverage option.">
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100 <validator type="regex" message="Genome size must be a float or integer, optionally followed by the a unit prefix (kmg)">^([0-9]*[.])?[0-9]+[kmg]?$</validator>
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101 </param>
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102 </when>
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103 <when value="false" />
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104 </conditional>
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105 <param argument="--no-alt-contigs" type="boolean" truevalue="--no-alt-contigs" falsevalue="" checked="false" label="Remove all non-primary contigs from the assembly"/>
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106 <param name="generate_log" type="boolean" truevalue="true" falsevalue="false" checked="false" label="Generate a log file"/>
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107 </inputs>
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108 <outputs>
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109 <data name="consensus" format="fasta" from_work_dir="out_dir/assembly.fasta" label="${tool.name} on ${on_string}: consensus"/>
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110 <data name="assembly_graph" format="graph_dot" from_work_dir="out_dir/assembly_graph.gv" label="${tool.name} on ${on_string}: assembly graph"/>
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111 <data name="assembly_gfa" format="gfa1" from_work_dir="out_dir/assembly_graph.gfa" label="${tool.name} on ${on_string}: graphical fragment assembly"/>
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112 <data name="assembly_info" format="tabular" from_work_dir="out_dir/assembly_info.txt" label="${tool.name} on ${on_string}: assembly info"/>
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113 <data name="flye_log" format="txt" from_work_dir="out_dir/flye.log" label="${tool.name} on ${on_string}: log">
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114 <filter>generate_log</filter>
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115 </data>
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116 </outputs>
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117 <tests>
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118 <!--Test 01: pacbio-raw-->
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119 <test expect_num_outputs="5">
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120 <param name="inputs" ftype="fastq.gz" value="ecoli_01.fastq.gz,ecoli_02.fastq.gz,ecoli_03.fastq.gz,ecoli_04.fastq.gz,ecoli_05.fastq.gz,ecoli_06.fastq.gz,ecoli_07.fastq.gz"/>
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121 <conditional name="mode_conditional">
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122 <param name="mode" value="--pacbio-raw"/>
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123 </conditional>
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124 <param name="iterations" value="0"/>
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125 <param name="generate_log" value="true"/>
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126 <output name="assembly_info" file="result1_assembly_info.txt" ftype="tabular" compare="sim_size"/>
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127 <output name="assembly_graph" file="result1_assembly_graph.dot" ftype="graph_dot" compare="sim_size"/>
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128 <output name="assembly_gfa" file="result1_assembly_graph.gfa" ftype="gfa1" compare="diff" lines_diff="10"/>
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129 <output name="consensus" ftype="fasta">
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130 <assert_contents>
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131 <has_line line=">contig_1"/>
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132 </assert_contents>
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133 </output>
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134 <output name="flye_log" file="result1.log" ftype="txt" compare="sim_size"/>
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135 </test>
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136 <!--Test 02: nano raw-->
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137 <test expect_num_outputs="4">
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138 <param name="inputs" ftype="fasta.gz" value="nanopore.fasta.gz"/>
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139 <conditional name="mode_conditional">
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140 <param name="mode" value="--nano-raw"/>
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141 </conditional>
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142 <param name="iterations" value="0"/>
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143 <output name="assembly_info" ftype="tabular">
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144 <assert_contents>
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145 <has_size value="95" delta="100"/>
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146 </assert_contents>
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147 </output>
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148 <output name="assembly_graph" ftype="graph_dot">
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149 <assert_contents>
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150 <has_size value="803" delta="100"/>
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151 </assert_contents>
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152 </output>
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153 <output name="assembly_gfa" ftype="gfa1">
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154 <assert_contents>
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155 <has_size value="35047" delta="100"/>
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156 </assert_contents>
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157 </output>
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158 <output name="consensus" ftype="fasta">
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159 <assert_contents>
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160 <has_size value="35573" delta="100"/>
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161 </assert_contents>
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162 </output>
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163 </test>
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164 <!--Test 03: reduce coverage-->
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165 <test expect_num_outputs="4">
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166 <param name="inputs" ftype="fastq.gz" value="ecoli_hifi_01.fastq.gz,ecoli_hifi_02.fastq.gz,ecoli_hifi_03.fastq.gz,ecoli_hifi_04.fastq.gz,ecoli_hifi_05.fastq.gz,ecoli_hifi_06.fastq.gz,ecoli_hifi_07.fastq.gz,ecoli_hifi_08.fastq.gz,ecoli_hifi_09.fastq.gz"/>
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167 <conditional name="mode_conditional">
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168 <param name="mode" value="--nano-raw"/>
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169 </conditional>
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170 <conditional name="asm">
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171 <param name="asm_select" value="true" />
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172 <param name="asm_coverage" value="30"/>
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173 <param name="genome_size" value="3980000"/>
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174 </conditional>
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175 <output name="assembly_info" ftype="tabular">
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176 <assert_contents>
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177 <has_size value="286" delta="100"/>
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178 </assert_contents>
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179 </output>
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180 <output name="assembly_graph" ftype="graph_dot">
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181 <assert_contents>
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182 <has_size value="1840" delta="100"/>
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183 </assert_contents>
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184 </output>
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185 <output name="assembly_gfa" ftype="gfa1">
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186 <assert_contents>
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187 <has_size value="420752" delta="100"/>
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188 </assert_contents>
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189 </output>
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190 <output name="consensus" ftype="fasta">
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191 <assert_contents>
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192 <has_size value="427580" delta="100"/>
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193 </assert_contents>
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194 </output>
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195 </test>
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196 <!--Test 04: metagenomic mode-->
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197 <test expect_num_outputs="4">
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198 <param name="inputs" ftype="fastq.gz" value="ecoli_01.fastq.gz,ecoli_02.fastq.gz,ecoli_03.fastq.gz,ecoli_04.fastq.gz,ecoli_05.fastq.gz,ecoli_06.fastq.gz,ecoli_07.fastq.gz"/>
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199 <conditional name="mode_conditional">
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200 <param name="mode" value="--pacbio-raw"/>
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201 </conditional>
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202 <param name="meta" value="true"/>
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203 <output name="assembly_info" ftype="tabular">
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204 <assert_contents>
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205 <has_size value="95" delta="100"/>
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206 </assert_contents>
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207 </output>
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208 <output name="assembly_graph" ftype="graph_dot">
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209 <assert_contents>
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210 <has_size value="367" delta="100"/>
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211 </assert_contents>
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212 </output>
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213 <output name="assembly_gfa" ftype="gfa1">
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214 <assert_contents>
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215 <has_size value="418729" delta="100"/>
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216 </assert_contents>
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217 </output>
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218 <output name="consensus" ftype="fasta">
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219 <assert_contents>
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220 <has_size value="425667" delta="100"/>
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221 </assert_contents>
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222 </output>
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223 </test>
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224 <!--Test 05: nanopore HQ mode-->
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225 <test expect_num_outputs="4">
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226 <param name="inputs" ftype="fastq.gz" value="ecoli_hifi_01.fastq.gz,ecoli_hifi_02.fastq.gz,ecoli_hifi_03.fastq.gz,ecoli_hifi_04.fastq.gz,ecoli_hifi_05.fastq.gz,ecoli_hifi_06.fastq.gz,ecoli_hifi_07.fastq.gz,ecoli_hifi_08.fastq.gz,ecoli_hifi_09.fastq.gz"/>
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227 <conditional name="mode_conditional">
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228 <param name="mode" value="--nano-hq"/>
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229 </conditional>
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230 <param name="min_overlap" value="1000"/>
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231 <output name="assembly_info" ftype="tabular">
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232 <assert_contents>
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233 <has_size value="286" delta="100"/>
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234 </assert_contents>
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235 </output>
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236 <output name="assembly_graph" ftype="graph_dot">
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237 <assert_contents>
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238 <has_size value="1248" delta="100"/>
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239 </assert_contents>
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240 </output>
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241 <output name="assembly_gfa" ftype="gfa1">
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242 <assert_contents>
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243 <has_size value="419414" delta="1000"/>
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244 </assert_contents>
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245 </output>
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246 <output name="consensus" ftype="fasta">
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247 <assert_contents>
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248 <has_size value="426277" delta="1000"/>
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249 </assert_contents>
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250 </output>
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251 </test>
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252 <!--Test 06: hifi error option-->
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253 <test expect_num_outputs="4">
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254 <param name="inputs" ftype="fastq.gz" value="ecoli_hifi_01.fastq.gz,ecoli_hifi_02.fastq.gz,ecoli_hifi_03.fastq.gz,ecoli_hifi_04.fastq.gz,ecoli_hifi_05.fastq.gz,ecoli_hifi_06.fastq.gz,ecoli_hifi_07.fastq.gz,ecoli_hifi_08.fastq.gz,ecoli_hifi_09.fastq.gz"/>
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255 <conditional name="mode_conditional">
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256 <param name="mode" value="--pacbio-hifi"/>
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257 <param name="hifi_error" value="0.21"/>
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258 </conditional>
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259 <param name="min_overlap" value="1000"/>
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260 <output name="assembly_info" ftype="tabular">
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261 <assert_contents>
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262 <has_size value="286" delta="100"/>
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263 </assert_contents>
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264 </output>
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265 <output name="assembly_graph" ftype="graph_dot">
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266 <assert_contents>
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267 <has_size value="1248" delta="500"/>
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268 </assert_contents>
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269 </output>
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270 <output name="assembly_gfa" ftype="gfa1">
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271 <assert_contents>
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272 <has_size value="420254" delta="2000"/>
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273 </assert_contents>
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274 </output>
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275 <output name="consensus" ftype="fasta">
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276 <assert_contents>
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277 <has_size value="427131" delta="2000"/>
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278 </assert_contents>
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279 </output>
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280 </test>
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281 <!--Test 07: keep haplotypes-->
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282 <test expect_num_outputs="4">
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283 <param name="inputs" ftype="fastq.gz" value="ecoli_hifi_01.fastq.gz,ecoli_hifi_02.fastq.gz,ecoli_hifi_03.fastq.gz,ecoli_hifi_04.fastq.gz,ecoli_hifi_05.fastq.gz,ecoli_hifi_06.fastq.gz,ecoli_hifi_07.fastq.gz,ecoli_hifi_08.fastq.gz,ecoli_hifi_09.fastq.gz"/>
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284 <conditional name="mode_conditional">
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285 <param name="mode" value="--pacbio-corr"/>
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286 </conditional>
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287 <param name="min_overlap" value="1000"/>
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288 <param name="keep_haplotypes" value="true"/>
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289 <output name="assembly_info" ftype="tabular">
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290 <assert_contents>
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291 <has_size value="286" delta="200"/>
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292 </assert_contents>
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293 </output>
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294 <output name="assembly_graph" ftype="graph_dot">
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295 <assert_contents>
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296 <has_size value="1273" delta="500"/>
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297 </assert_contents>
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298 </output>
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299 <output name="assembly_gfa" ftype="gfa1">
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300 <assert_contents>
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301 <has_size value="420254" delta="3000"/>
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302 </assert_contents>
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303 </output>
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304 <output name="consensus" ftype="fasta">
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305 <assert_contents>
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306 <has_size value="427131" delta="3000"/>
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307 </assert_contents>
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308 </output>
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309 </test>
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310 <!--Test 08: scaffolding mode-->
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311 <test expect_num_outputs="4">
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312 <param name="inputs" ftype="fastq.gz" value="ecoli_hifi_01.fastq.gz,ecoli_hifi_02.fastq.gz,ecoli_hifi_03.fastq.gz,ecoli_hifi_04.fastq.gz,ecoli_hifi_05.fastq.gz,ecoli_hifi_06.fastq.gz,ecoli_hifi_07.fastq.gz,ecoli_hifi_08.fastq.gz,ecoli_hifi_09.fastq.gz"/>
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313 <conditional name="mode_conditional">
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314 <param name="mode" value="--nano-hq"/>
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315 </conditional>
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316 <param name="min_overlap" value="1000"/>
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317 <param name="scaffold" value="true"/>
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318 <output name="assembly_info" ftype="tabular">
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319 <assert_contents>
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320 <has_size value="286" delta="100"/>
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321 </assert_contents>
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322 </output>
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323 <output name="assembly_graph" ftype="graph_dot">
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324 <assert_contents>
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325 <has_size value="1248" delta="100"/>
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326 </assert_contents>
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327 </output>
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328 <output name="assembly_gfa" ftype="gfa1">
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329 <assert_contents>
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330 <has_size value="419414" delta="2000"/>
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331 </assert_contents>
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332 </output>
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333 <output name="consensus" ftype="fasta">
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334 <assert_contents>
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335 <has_size value="426277" delta="2000"/>
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336 </assert_contents>
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337 </output>
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338 </test>
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339 <!--Test 09: test not-alt-contigs parameter w-->
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340 <test expect_num_outputs="4">
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341 <param name="inputs" ftype="fasta.gz" value="nanopore.fasta.gz"/>
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342 <conditional name="mode_conditional">
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343 <param name="mode" value="--nano-raw"/>
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344 </conditional>
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345 <param name="iterations" value="0"/>
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346 <param name="no_alt_contigs" value="true"/>
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347 <output name="assembly_info" ftype="tabular">
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348 <assert_contents>
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349 <has_size value="151" delta="100"/>
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350 </assert_contents>
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351 </output>
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352 <output name="assembly_graph" ftype="graph_dot">
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353 <assert_contents>
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354 <has_size value="217" delta="100"/>
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355 </assert_contents>
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356 </output>
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357 <output name="assembly_gfa" ftype="gfa1">
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358 <assert_contents>
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359 <has_size value="5110" delta="100"/>
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360 </assert_contents>
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361 </output>
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362 <output name="consensus" ftype="fasta">
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363 <assert_contents>
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364 <has_size value="5123" delta="100"/>
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365 </assert_contents>
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366 </output>
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367 </test>
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368 </tests>
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369 <help><![CDATA[
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370
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371 **Purpose**
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372
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373 Flye is a de novo assembler for single molecule sequencing reads, such as those produced by PacBio and Oxford Nanopore Technologies.
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374 It is designed for a wide range of datasets, from small bacterial projects to large mammalian-scale assemblies. The package represents
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375 a complete pipeline: it takes raw PacBio/ONT reads as input and outputs polished contigs. Flye also has a special mode for metagenome
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376 assembly.
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377
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378 ----
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379
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380 **Quick usage**
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381
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382 Input reads can be in FASTA or FASTQ format, uncompressed or compressed with gz. Currently, PacBio (raw, corrected, HiFi) and ONT reads
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383 (raw, corrected) are supported. Expected error rates are <30% for raw, <3% for corrected, and <1% for HiFi. Note that Flye was primarily
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384 developed to run on raw reads. You may specify multiple files with reads (separated by spaces). Mixing different read types is not yet supported. The *--meta* o
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385 ption enables the mode for metagenome/uneven coverage assembly.
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386
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387 Genome size estimate is no longer a required option. You need to provide an estimate if using *--asm-coverage* option.
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388
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389 To reduce memory consumption for large genome assemblies, you can use a subset of the longest reads for initial disjointig assembly by
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390 specifying *--asm-coverage* and *--genome-size* options. Typically, 40x coverage is enough to produce good disjointigs.
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391
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392 ----
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393
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394 **Outputs**
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395
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396 The main output files are:
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397
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398 * Final assembly: contains contigs and possibly scaffolds (see below).
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399 * Final repeat graph: note that the edge sequences might be different (shorter) than contig sequences, because contigs might include multiple graph edges.
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400 * Extra information about contigs (such as length or coverage).
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401
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402 Each contig is formed by a single unique graph edge. If possible, unique contigs are extended with the sequence from flanking unresolved repeats on the graph. Thus,
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403 a contig fully contains the corresponding graph edge (with the same id), but might be longer then this edge. This is somewhat similar to unitig-contig relation in
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404 OLC assemblers. In a rare case when a repetitive graph edge is not covered by the set of "extended" contigs, it will be also output in the assembly file.
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405
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406 Sometimes it is possible to further order contigs into scaffolds based on the repeat graph structure. These ordered contigs will be output as a part of scaffold in
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407 the assembly file (with a scaffold prefix). Since it is hard to give a reliable estimate of the gap size, those gaps are represented with the default 100 Ns.
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408 assembly_info.txt file (below) contains additional information about how scaffolds were formed.
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409
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410 Extra information about contigs/scaffolds is output into the assembly_info.txt file. It is a tab-delimited table with the columns as follows:
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411
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412 * Contig/scaffold id
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413 * Length
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414 * Coverage
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415 * Is circular, (Y)es or (N)o
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416 * Is repetitive, (Y)es or (N)o
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417 * Multiplicity (based on coverage)
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418 * Alternative group
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419 * Graph path (graph path corresponding to this contig/scaffold).
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420
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421 Scaffold gaps are marked with `??` symbols, and `*` symbol denotes a terminal graph node. Alternative contigs (representing alternative haplotypes) will have the same alt.
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422 group ID. Primary contigs are marked by `*`.
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423
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424 ----
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425
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426 **Algorithm Description**
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427
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428 This is a brief description of the Flye algorithm. Please refer to the manuscript for more detailed information. The draft contig extension is organized as follows:
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429
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430 * K-mer counting / erroneous k-mer pre-filtering
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431 * Solid k-mer selection (k-mers with sufficient frequency, which are unlikely to be erroneous)
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432 * Contig extension. The algorithm starts from a single read and extends it with a next overlapping read (overlaps are dynamically detected using the selected solid k-mers).
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433
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434 Note that we do not attempt to resolve repeats at this stage, thus the reconstructed contigs might contain misassemblies. Flye then aligns the reads on these draft
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435 contigs using minimap2 and calls a consensus. Afterwards, Flye performs repeat analysis as follows:
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436
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437 * Repeat graph is constructed from the (possibly misassembled) contigs
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438 * In this graph all repeats longer than minimum overlap are collapsed
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439 * The algorithm resolves repeats using the read information and graph structure
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440 * The unbranching paths in the graph are output as contigs
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441
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442 If enabled, after resolving bridged repeats, Trestle module attempts to resolve simple unbridged repeats (of multiplicity 2) using the heterogeneities between repeat copies.
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443 Finally, Flye performs polishing of the resulting assembly to correct the remaining errors:
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444
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445 * Alignment of all reads to the current assembly using minimap2
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446 * Partition the alignment into mini-alignments (bubbles)
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447 * Error correction of each bubble using a maximum likelihood approach
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448
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449 The polishing steps could be repeated, which might slightly increase quality for some datasets.
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450
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451
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452 ]]></help>
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453 <expand macro="citations" />
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454 </tool>