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1 <tool id="Readmap" name="Generate readmap and histograms from alignment files" version="1.0.0">
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2 <description>from sRbowtie aligment</description>
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3 <requirements>
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4 <requirement type="package" version="0.12.7">bowtie</requirement>
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5 <requirement type="package" version="0.1.18">samtools</requirement>
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6 <requirement type="package" version="0.7.7">pysam</requirement>
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7 <requirement type="package" version="2.14">biocbasics</requirement>
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8 <requirement type="package" version="3.0.3">R</requirement>
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9 </requirements>
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10 <parallelism method="basic"></parallelism>
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11 <command interpreter="python">
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12 readmap.py
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13 #if $refGenomeSource.genomeSource == "history":
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14 --reference_fasta ## sys.argv[2]
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15 $refGenomeSource.ownFile ## index source
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16 #else:
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17 #silent reference= filter( lambda x: str( x[0] ) == str( $refGenomeSource.series[0].input.dbkey ), $__app__.tool_data_tables[ 'bowtie_indexes' ].get_fields() )[0][-1]
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18 --reference_bowtie_index
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19 $reference
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20 #end if
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21 --rcode
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22 $plotCode
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23 --output_readmap
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24 $readmap_dataframe
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25 --output_size_distribution
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26 $size_distribution_dataframe
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27 --minquery
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28 $minquery
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29 --maxquery
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30 $maxquery
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31 --input
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32 #for $i in $refGenomeSource.series
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33 $i.input
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34 #end for
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35 --ext
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36 #for $i in $refGenomeSource.series
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37 $i.input.ext
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38 #end for
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39 --label
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40 #for $i in $refGenomeSource.series
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41 "$i.input.name"
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42 #end for
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43 --normalization_factor
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44 #for $i in $refGenomeSource.series
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45 $i.norm
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46 #end for
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47 #if $gff:
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48 --gff
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49 $gff
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50 #end if
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51
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52 </command>
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53 <inputs>
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54 <conditional name="refGenomeSource">
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55 <param name="genomeSource" 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">
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56 <option value="indexed">Use a built-in index</option>
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57 <option value="history">Use one from the history</option>
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58 </param>
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59 <when value="indexed">
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60 <repeat name="series" title="Add alignment files">
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61 <param name="input" type="data" label="Select multiple alignments to parse">
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62 <validator type="dataset_metadata_in_data_table" table_name="bowtie_indexes" metadata_name="dbkey" metadata_column="0" message="database not set for this bowtie output. Select the database(=genome used for matching) manually, or select a reference fasta from your history."/>
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63 </param>
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64 <param name="norm" type="float" value="1" label="Indicate a normalization factor to compare multiple aligments"/>
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65 </repeat>
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66 </when>
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67 <when value="history">
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68 <param name="ownFile" type="data" format="fasta" label="Select a fasta file, that served as the reference index for the alignments" />
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69 <repeat name="series" title="Add alignment files">
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70 <param name="input" type="data" label="Select multiple alignments to parse"/>
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71 <param name="norm" type="float" value="1" label="Indicate a normalization factor to compare multiple aligments"/>
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72 </repeat>
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73 </when>
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74 </conditional>
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75 <param name="gff" type="data" format="gff3" optional="true" label="Optional: select a GFF to investigate regions of interest" help="GFF must match genome build"/>
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76 <!-- <validator type="dataset_metadata_in_data_table" table_name="bowtie_indexes" metadata_name="dbkey" metadata_column="0" message="GFF database and alignment file databse do not match!"/> -->
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77 <param name="minquery" type="integer" size="3" value="18" label="Min size of query small RNAs" help="'18' = 18 nucleotides"/>
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78 <param name="maxquery" type="integer" size="3" value="28" label="Max size of query small RNAs" help="'28' = 28 nucleotides"/>
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79 <param name="title" type="text" size="15" value= "Readmaps and size distributions" label="Main Titles"/>
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80 <param name="xlabel" type="text" size="15" value="Coordinates/read size" label="x axis label"/>
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81 <param name="ylabel" type="text" size="15" value="Number of reads" label="y axis label"/>
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82 <param name="rows_per_page" type="text" size="9" value="8" label="How many items to display per page?">
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83 <validator type="in_range" min="6" max="20" message="Select between 6 and 20 rows, as the readability will suffer otherwise."/>
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84 </param>
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85 </inputs>
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86 <configfiles>
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87 <configfile name="plotCode">
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88 ## Setup R error handling to go to stderr
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89 options( show.error.messages=F,
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90 error = function () { cat( geterrmessage(), file=stderr() ); q( "no", 1, F ) } )
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91 library(RColorBrewer)
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92 library(lattice)
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93 library(latticeExtra)
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94 library(grid)
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95 library(gridExtra)
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96
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97 ## data frames implementation
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98
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99 rm=read.delim("${readmap_dataframe}", header=T, row.names=NULL)
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100 n_samples=length(unique(rm\$sample))
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101 genes=unique(levels(rm\$gene))
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102 per_gene_readmap=lapply(genes, function(x) subset(rm, gene==x)) ####### ?
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103 n_genes=length(per_gene_readmap)
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104
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105 size=read.delim("${size_distribution_dataframe}", header=T, row.names=NULL)
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106 per_gene_size=lapply(genes, function(x) subset(size, gene==x)) ###### ?
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107
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108 ## end of data frames implementation
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109
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110 ## functions
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111
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112 plot_readmap=function(df, ...) {
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113 combineLimits(xyplot(count~coord|factor(sample, levels=unique(sample))+reorder(gene, count, function(x) -sum(abs(x))),
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114 data=df,
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115 type='h',
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116 scales= list(relation="free", x=list(rot=0, cex=0.7, axs="i", tck=0.5), y=list(tick.number=4, rot=90, cex=0.7)),
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117 xlab=NULL, main=NULL, ylab=NULL,
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118 as.table=T,
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119 origin = 0,
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120 horizontal=FALSE,
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121 group=polarity,
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122 col=c("red","blue"),
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123 par.strip.text = list(cex=0.7),
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124 ...))
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125 }
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126
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127 plot_size_distribution= function(df, ...) {
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128 smR.prepanel=function(x,y,...){; yscale=c(-max(abs(y)), max(abs(y)));list(ylim=yscale);}
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129 bc= barchart(count~as.factor(size)|factor(sample, levels=unique(sample))+gene, data = df, origin = 0,
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130 horizontal=FALSE,
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131 group=polarity,
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132 stack=TRUE,
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133 col=c('red', 'blue'),
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134 cex=0.75,
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135 scales=list(y=list(tick.number=4, rot=90, relation="free", cex=0.7), x=list(cex=0.7) ),
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136 prepanel=smR.prepanel,
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137 xlab = NULL,
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138 ylab = NULL,
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139 main = NULL,
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140 as.table=TRUE,
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141 newpage = T,
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142 par.strip.text = list(cex=0.7),
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143 ...)
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144 combineLimits(bc)
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145 }
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146
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147 ## end of functions
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148
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149 ## function parameters'
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150
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151 par.settings.readmap=list(layout.heights=list(top.padding=0, bottom.padding=-2.5), strip.background = list(col=c("lightblue","lightgreen")) )
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152 par.settings.size=list(layout.heights=list(top.padding=-1, bottom.padding=-2.5), strip.background = list(col=c("lightblue","lightgreen")) )
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153 par.settings.combination.readmap=list(layout.heights=list(top.padding=0, bottom.padding=-3), strip.background=list(col=c("lightblue","lightgreen")) )
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154 par.settings.combination.size=list(layout.heights=list(top.padding=-2, bottom.padding=-0.5), strip.background=list(col=c("lightblue", "lightgreen")) )
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155
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156 ## end of function parameters'
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157
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158 ## GRAPHS
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159
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160 if (n_genes > 7) {page_height_simple = 11.69; page_height_combi=11.69; rows_per_page=${rows_per_page}; extrarow=0 } else {
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161 rows_per_page= n_genes; page_height_simple = 11.69/n_genes/4; page_height_combi=11.69/(n_genes*2); extrarow=1 }
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162 if (n_samples > 4) {page_width = 8.2677*n_samples/4} else {page_width = 8.2677*n_samples/3} # to test
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163
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164 pdf(file="${readmap_PDF}", paper="special", height=page_height_simple, width=page_width)
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165 for (i in seq(1,n_genes,rows_per_page)) {
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166 start=i
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167 end=i+rows_per_page-1
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168 if (end>n_genes) {end=n_genes}
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169 readmap_plot.list=lapply(per_gene_readmap[start:end], function(x) plot_readmap(x, par.settings=par.settings.readmap))
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170 args.list=c(readmap_plot.list, list(nrow=rows_per_page, ncol=1,
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171 main=textGrob("Read Maps (nucleotide coordinates)", gp=gpar(cex=1), just="top"),
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172 left=textGrob("${ylabel}", gp=gpar(cex=1), vjust=1, rot=90)
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173 #sub=textGrob("readmap coordinates", gp=gpar(cex=.75), just="bottom")
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174 )
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175 )
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176 do.call(grid.arrange, args.list)
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177 }
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178 devname=dev.off()
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179
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180
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181 pdf(file="${size_PDF}", paper="special", height=page_height_simple, width=page_width)
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182 for (i in seq(1,n_genes,rows_per_page)) {
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183 start=i
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184 end=i+rows_per_page-1
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185 if (end>n_genes) {end=n_genes}
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186 plot.list=lapply(per_gene_size[start:end], function(x) plot_size_distribution(x, par.settings=par.settings.size) )
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187 args.list=c(plot.list, list(nrow=rows_per_page, ncol=1,
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188 main=textGrob("Size distributions (in nucleotides)", gp=gpar(cex=1), just="top"),
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189 left=textGrob("${ylabel}", gp=gpar(cex=1), vjust=1, rot=90)
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190 #sub="readsize in nucleotides"
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191 )
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192 )
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193 do.call(grid.arrange, args.list)
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194 }
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195 devname=dev.off()
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196
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197 pdf(file="${combi_PDF}", paper="special", height=page_height_combi, width=page_width)
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198 for (i in seq(1,n_genes,rows_per_page/2)) {
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199 start=i
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200 end=i+rows_per_page/2-1
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201 if (end>n_genes) {end=n_genes}
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202 read_plot.list=lapply(per_gene_readmap[start:end], function(x) plot_readmap(x, par.settings=par.settings.combination.readmap))
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203 size_plot.list=lapply(per_gene_size[start:end], function(x) plot_size_distribution(x, strip=FALSE, par.settings=par.settings.combination.size))
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204 plot.list=rbind(read_plot.list, size_plot.list )
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205 args.list=c(plot.list, list(nrow=rows_per_page + extrarow, ncol=1,
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206 main=textGrob("${title}", gp=gpar(cex=1), just="top"),
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207 left=textGrob("${ylabel}", gp=gpar(cex=1), vjust=1, rot=90),
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208 sub=textGrob("${xlabel}", gp=gpar(cex=1), just="bottom")
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209 )
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210 )
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211 do.call(grid.arrange, args.list)
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212 }
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213 devname=dev.off()
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214
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215
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216 </configfile>
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217 </configfiles>
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218
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219 <outputs>
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220 <data format="tabular" name="readmap_dataframe" label="Readmap dataframe"/>
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221 <data format="tabular" name="size_distribution_dataframe" label="Size distribution dataframe"/>
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222 <data format="pdf" name="readmap_PDF" label="Readmaps"/>
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223 <data format="pdf" name="size_PDF" label="Size distribution"/>
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224 <data format="pdf" name="combi_PDF" label="Size distribution and Readmaps"/>
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225 </outputs>
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226 <help>
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227
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228 **What it does**
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229
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230 Takes one or more alignment files (BAM, SAM or tabular bowtie output) as input and produces a "Readmap",
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231 where by default for each "chromosome" the position of the read is recorded on the x-axis, and the y-axis indicates
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232 the number of reads per position. Reads that map in sense are on the top, reads that map antisense are on the bottom.
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233
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234
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235 .. class:: warningmark
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236
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237 '''TIP''' The input data can be produced using the sRbowtie tool.
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238
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239 ----
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240
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241 '''Example'''
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242
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243 Query sequence::
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244 For a SAM file as the following:
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245
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246 5 16 2L_79 24393 255 17M * 0 0 CCTTCATCTTTTTTTTT IIIIIIIIIIIIIIIII XA:i:0 MD:Z:17 NM:i:0
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247
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248 11 0 2R_1 12675 255 21M * 0 0 AAAAAAAACGCGTCCTTGTGC IIIIIIIIIIIIIIIIIIIII XA:i:0 MD:Z:21 NM:i:0
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249
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250 2 16 2L_5 669 255 23M * 0 0 TGTTGCTGCATTTCTTTTTTTTT IIIIIIIIIIIIIIIIIIIIIII XA:i:0 MD:Z:23 NM:i:0
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251
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252 produce a plot like this:
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253
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254 ----
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255
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256 .. image:: static/images/readmap.png
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257 :height: 800
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258 :width: 500
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259
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260 </help>
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261
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262 <test>
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263 <param name="genomeSource" value="history" />
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264 <param name="ownFile" value ="transposons.fasta" ftype="fasta" />
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265 <param name="input" value="sample1.srbowtie_out, sample2.srbowtie_out, sample3.srbowtie_out" ftype="tabular" />
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266 <param name="norm" value="1,1,1" />
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267 <param name="minquery" value="20" />
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268 <param name="maxquery" value="30" />
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269 <param name="title" value="Readmaps and size distributions" />
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270 <param name="xlabel" value="Coordinates/read size" />
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271 <param name="ylabel" value="Number of reads" />
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272 <param name="rows_per_page" value="8" />
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273 <output name="readmap_dataframe" ftype="tabular" value="Readmap_dataframe.tab" />
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274 <output name="size_distribution_dataframe" ftype="tabular" value="Size_distribution_dataframe.tab" />
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275 <output name="readmap_PDF" ftype="pdf" value="Readmaps.pdf" />
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276 <output name="size_PDF" ftype="pdf" value="Size_distribution.pdf" />
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277 <output name="combi_PDF" ftype="pdf" value="Size_distribution_and_Readmaps.pdf" />
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278 </test>
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279
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280 </tool>
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