annotate umi-tools_counts.xml @ 0:8db56d2f8b72 draft

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date Thu, 21 Jun 2018 15:20:14 -0400
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1 <tool id="umi_tools_count" name="UMI-tools count" version="@VERSION@.0">
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2 <description>Count UMIs from BAM files</description>
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3 <macros>
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4 <import>macros.xml</import>
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5 <xml name="sanitize_tag" >
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6 <sanitizer invalid_char="">
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7 <valid initial="string.letters,string.digits" />
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8 </sanitizer>
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9 </xml>
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10 </macros>
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11 <expand macro="requirements" />
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12 <command detect_errors="exit_code"><![CDATA[
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13
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14 ln -s '${input_bam}' 'input.bam' &&
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15 ln -s '${input_bam.metadata.bam_index}' 'input.bam.bai' &&
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16
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17 umi_tools count
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18 -I input.bam
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19 '$bam_paired'
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20 --extract-umi-method='$barcodes.extract_umi_method.value'
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21 #if $barcodes.extract_umi_method == 'read_id':
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22 --umi-separator='$barcodes.delimiter'
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23 #else if $barcodes.extract_umi_method == 'tag':
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24 --umi-tag='$barcodes.umi_tag'
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25 --cell-tag='$barcodes.cell_tag'
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26 #end if
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27 --method='$grouping_method.value'
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28 --edit-distance-threshold='$hamming_distance'
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29 --mapping-quality='$advanced.mapping_quality'
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30 --per-gene
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31 $wide_format_cell_counts
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32 $advanced.per_contig
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33 '$advanced.per_cell'
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34 #if $advanced.gene_tag:
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35 --gene-tag='$advanced.gene_tag'
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36 #end if
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37 #if $advanced.skip_tags_regex.value:
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38 --skip-tags-regex='$advanced.skip_tags_regex'
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39 #end if
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40 #if $advanced.random_seed != 0:
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41 --random-seed='$advanced.random_seed'
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42 #end if
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43 -S '$out_counts'
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44 -L '$out_log'
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45 ]]></command>
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46 <inputs>
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47 <param name="input_bam" type="data" format="bam" label="Sorted BAM file" help="Please use the samtools sort tool to ensure a correct BAM input" />
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48
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49 <param name="bam_paired" type="boolean" truevalue="--paired" falsevalue="" checked="false"
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50 label="Bam is paired-end"
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51 help="both read pairs will be output. This will also force the use of the template length to determine
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52 reads with the same mapping coordinates." />
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53
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54 <conditional name="barcodes" >
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55 <param name="extract_umi_method" type="select" label="Umi Extract Method" help="How are the barcodes encoded in the read?" >
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56 <option value="read_id" selected="true">Barcodes are contained at the end of the read seperated by a delimiter</option>
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57 <option value="tag" >Barcodes are contained in tags</option>
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58 <option value="umis" >Barcodes were extracted using umis</option>
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59 </param>
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60 <when value="read_id" >
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61 <param name="delimiter" type="text" label="Delimiter between read id and the UMI" value="_" >
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62 <expand macro="sanitize_tag" />
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63 </param>
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64 </when>
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65 <when value="tag" >
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66 <param name="umi_tag" type="text" label="Tag which contains the UMI" >
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67 <expand macro="sanitize_tag" />
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68 </param>
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69 <param name="cell_tag" type="text" label="Tag which contains the cell barcode" >
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70 <expand macro="sanitize_tag" />
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71 </param>
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72 </when>
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73 <when value="umis"></when>
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74 </conditional>
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75
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76 <param name="grouping_method" type="select" label="Method to identify group of reads" help="UMIs with the same (or similar) codes can be grouped together. The simplest methods 'unique' and 'percentile' group identical
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77 UMIs, however 'cluster', 'adjacency', and 'directional' can group similar umis with edit distances less than some threshold. Unique: Reads group share the exact same UMI. Percentile: Reads group share the same UMI, and UMIs with
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78 counts &lt; 1% of the median counts for UMIs at the same position are ignored. Cluster: Identify clusters of connected UMIs (based on hamming distance threshold). Adjacency: Same as cluster, but considers only directly ajacent
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79 UMIs in the cluster. Directional: Identify cluster of connected UMIs based on hamming distance and umi." >
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80 <option value="unique" >Unique</option>
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81 <option value="percentile">Percentile</option>
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82 <option value="cluster">Cluster</option>
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83 <option value="adjacency">Adjacency</option>
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84 <option value="directional" selected="true" >Directional</option>
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85 </param>
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86
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87 <param name="hamming_distance" type="integer" label="Edit distance threshold" min="0" value="1" />
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88 <param name="wide_format_cell_counts" type="boolean" truevalue="--wide-format-cell-counts" falsevalue="" checked="false" label="Output a mtrix of genes and cells, instead of a flat file" />
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89
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90 <section name="advanced" title="Extra parameters" >
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91 <param name="mapping_quality" type="integer" min="0" value="0" label="Minimum mapping quality" />
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92 <!-- Currently hard-coded parameter. Leave here if useful to future wrapper -->
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93 <!-- <param argument="-\-per-gene" name="per_gene" type="text" label="Group reads together if they have the same gene" help="Reads will be grouped together if they have the same gene. This is useful if your library
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94 prep generates PCR duplicates with non-identical alignment positions such as CEL-Seq. Note this option is hardcoded to be on with the count command. I.e counting is always performed per-gene. Must be combined with either
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95 -\-gene-tag or -\-per-contig option" /> -->
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96 <param name="gene_tag" type="text" label="Deduplicate per gene." help="The gene information is encoded in the bam read tag." value="" >
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97 <expand macro="sanitize_tag" />
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98 </param>
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99 <param name="skip_tags_regex" type="text" label="Skip any reads where the gene matches this tag" value="" >
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100 <sanitizer invalid_char="">
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101 <valid initial="string.letters,string.digits">
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102 <add value="!="/>
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103 <add value="-"/>
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104 <add value="_"/>
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105 <add value="."/>
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106 <add value="?"/>
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107 <add value="&lt;"/><!-- left triangle bracket -->
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108 <add value="&gt;"/><!-- right triangle bracket -->
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109 <add value="&#91;"/> <!-- left square bracket -->
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110 <add value="&#93;"/> <!-- right square bracket -->
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111 <add value="&#94;"/> <!-- caret -->
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112 <add value="&#123;"/> <!-- left curly -->
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113 <add value="&#125;"/> <!-- right curly -->
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114 <add value="&#40;"/> <!-- left parenthesis -->
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115 <add value="&#41;"/> <!-- right parenthesis -->
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116 </valid>
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117 </sanitizer>
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118 </param>
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119 <param name="per_contig" type="boolean" truevalue="--per-contig" falsevalue="" checked="false"
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120 label="Deduplicate per contig (field 3 in BAM; RNAME)"
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121 help="All reads with the same contig will be considered to have the same alignment position. This is useful if you have aligned to a reference transcriptome with one transcript per gene." />
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122 <param name="per_cell" type="boolean" truevalue="--per-cell" falsevalue="" checked="false"
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123 label="Group reads only if they have the same cell barcode." />
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124 <param name="random_seed" type="integer" min="0" value="0" label="Random Seed" />
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125 </section>
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126 </inputs>
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127 <outputs>
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128 <data name="out_counts" format="tsv" />
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129 <data name="out_log" format="txt" />
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130 </outputs>
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131 <tests>
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132 <test><!--count_single_gene_tag:-->
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133 <param name="input_bam" value="chr19_gene_tags.bam" />
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134 <param name="random_seed" value="123456789" />
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135 <param name="grouping_method" value="directional" />
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136 <param name="gene_tag" value="XF" />
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137 <param name="skip_tags_regex" value="^[__|Unassigned]" />
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138 <param name="extract_umi_method" value="umis" />
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139 <output name="out_counts" value="count_single_gene_tag.tsv" />
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140 </test>
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141 <test><!--count_single_cells_gene_tag:-->
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142 <param name="input_bam" value="chr19_gene_tags.bam" />
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143 <param name="random_seed" value="123456789" />
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144 <param name="grouping_method" value="directional" />
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145 <param name="gene_tag" value="XF" />
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146 <param name="skip_tags_regex" value="^[__|Unassigned]" />
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147 <param name="per_cell" value="true" /><!-- new -->
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148 <param name="extract_umi_method" value="umis" />
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149 <output name="out_counts" value="count_single_cells_gene_tag.tsv" />
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150 </test>
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151 <test><!--count_single_cells_wide_gene_tag:-->
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152 <param name="input_bam" value="chr19_gene_tags.bam" />
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153 <param name="random_seed" value="123456789" />
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154 <param name="grouping_method" value="directional" />
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155 <param name="gene_tag" value="XF" />
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156 <param name="skip_tags_regex" value="^[__|Unassigned]" />
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157 <param name="per_cell" value="true" /><!-- new -->
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158 <param name="extract_umi_method" value="umis" />
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159 <param name="wide_format_cell_counts" value="true" />
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160 <output name="out_counts" value="count_single_cells_gene_tag_wide.tsv" />
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161 </test>
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162 </tests>
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163 <help><![CDATA[
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164
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165 UMI Tools count - Count reads per gene from BAM using UMIs
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166 ----------------------------------------------------------
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167
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168 Purpose
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169 -------
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170
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171 The purpose of this command is to count the number of reads per gene based
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172 on the mapping co-ordinate and the UMI attached to the read.
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173
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174
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175 It is assumed that the FASTQ files were processed with extract_umi.py
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176 before mapping and thus the UMI is the last word of the read name. e.g:
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177
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178 @HISEQ:87:00000000_AATT
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179
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180 where AATT is the UMI sequeuence.
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181
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182 If you have used an alternative method which does not separate the
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183 read id and UMI with a "_", such as bcl2fastq which uses ":", you can
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184 specify the separator, or if your UMIs are encoded in a tag you can also specify this.
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185
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186 ]]></help>
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187 <expand macro="citations" />
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188 </tool>