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1 #!/usr/bin/env Rscript
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2 # rm(list=ls())
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3
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4 #Rscript my_VDM_tool.R --inf "nor22.vcf" --itype "C.elegans" --qual 200 --allr "AB" --snp TRUE --lsp 0.4 --pcol "black" --lcol "red" --xstand TRUE --bsize 1000000 --freqthr 0.0-1.0 --bnorm FALSE --exclf "FALSE" --exclcol "green" --outn "test-output.txt" --pdfn "test-plot.pdf"
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5
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6 library("getopt")
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7 #input from trailing line arguments
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8 args <- commandArgs(trailingOnly = TRUE)
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9 #read the options from input commandArgs
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10 option_specification = matrix(c(
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11 'inf','in',1,'character',
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12 'itype','i',2,'character',
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13 'qual','q',2,'double',
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14 'allr','a',2,'character',
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15 'snp','s',2,'logical',
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16 'lsp','l',2,'double',
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17 'pcol','pc',2,'character',
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18 'lcol','lc',2,'character',
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19 'xstand','x',2,'logical',
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20 'bsize','b',2,'integer',
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21 'freqthr','ft',2,'character',
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22 'bnorm','n',2,'logical',
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23 'exclf','ef',2,'character',
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24 'exclcol','ec',2,'character',
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25 'outn','o',2,'character',
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26 'pdfn','p',2,'character'
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27 ), byrow=TRUE, ncol=4)
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28 options = getopt(option_specification)
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29
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30 myfunction<-function(inf,itype,qual,allr,snp,lsp,pcol,lcol,xstand,bsize,freqthr,bnorm,exclf,exclcol,outn,pdfn){
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31 #input file
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32 filename<-inf
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33 #choose species for chrom config
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34 interval_type<-itype
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35 #quality filter
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36 read_qual<-as.numeric(qual)
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37 #allele ratio from AB or AO/(AO+RO)
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38 allele_ratio<-allr
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39 #snp only or include all variant types
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40 snp_only<-snp
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41 #loess span
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42 loess_span<-as.numeric(lsp)
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43 #scatterplot point colour
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44 plot_color<-pcol
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45 #loess curve colour
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46 loess_color<-lcol
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47 #uniform y-axis scaling
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48 xaxis_standard<-xstand
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49 #binsize
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50 bin_size<-as.numeric(bsize)
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51
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52 #allele ratio greater or equal to this value considered hom ALT
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53 threshold_upper<-as.numeric(gsub(".*-","",freqthr))
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54 #allele ratio less than or equal to this value considered hom REF
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55 threshold_lower<-as.numeric(gsub("-.*","",freqthr))
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56
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57 #normalize frequency barplot
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58 bfreq_norm<-bnorm
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59 #filenames for variant subtraction
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60 exclusion_list<-exclf
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61 #pre-subtraction loess curve colour
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62 excl_loess_color<-exclcol
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63 #filename for ouput table
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64 vcfoutput_filename<-outn
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65 #filename for pdf
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66 pdf_filename<-pdfn
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67
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68 #transparency for selected colour (to see plot points underneath)
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69 plot_color<-rgb(c(col2rgb(plot_color)[1]),c(col2rgb(plot_color)[2]),c(col2rgb(plot_color)[3]),max=255,alpha=150)
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70 loess_color<-rgb(c(col2rgb(loess_color)[1]),c(col2rgb(loess_color)[2]),c(col2rgb(loess_color)[3]),max=255,alpha=150)
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71 excl_loess_color<-rgb(c(col2rgb(excl_loess_color)[1]),c(col2rgb(excl_loess_color)[2]),c(col2rgb(excl_loess_color)[3]),max=255,alpha=150)
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72
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73 ###FIXED PARAMETERS
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74 #chromosome intervals in Mb rather than custom
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75 interval_unit<-1000000
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76 #linkage scatter plot yaxis max value=1
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77 sp_yaxis<-1
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78
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79 #only use variants with allele ratio in this range for subtraction (dependent on AB/ratio option)
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80 excl_rthreshold_upper<-1
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81 excl_rthreshold_lower<-0
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82 #filename for parsed original vcf
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83 vcfparsed_filename<-"parsed.txt"
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84
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85 fastmode<-"TRUE"
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86
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87 ######################
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88 ###READ IN VCF FILE
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89 #extract column names
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90 vcf_readin<-readLines(filename)
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91 #find header line, i.e. last line to begin with #
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92 for(l in 1:length(vcf_readin)){
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93 vcf_readinl<-vcf_readin[l]
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94 if(substr(vcf_readinl,1,1)=="#"){next}
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95 else if(substr(vcf_readinl,1,1)!="#"){rowline<-l-1;break}
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96 }
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97 vcf_header<-vcf_readin[rowline]
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98 #e.g. CHROM\tPOS\tID\tREF\tALT\tQUAL\tFILTER\tINFO\tFORMAT\trgSM"
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99 vcf_header<-gsub("#","",vcf_header)
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100 vcf_colnames<-unlist(strsplit(vcf_header,"\t"))
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101
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102 #extract data (hashed vcf header skipped with read.table)
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103 vcf_rtable<-read.table(filename,sep="\t",stringsAsFactors=FALSE)
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104 names(vcf_rtable)<-vcf_colnames
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105
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106 if(fastmode=="TRUE"){
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107 #to speed up runtime- quality filter here ny skipping parsing of full vcf (parsed output file incomplete)
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108 vcf_rtable<-subset(vcf_rtable,QUAL>=read_qual)
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109 }
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110
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111
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112 ######################
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113 ###PREPARE DATA
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114 vcfinfo_dat<-NULL
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115 multiallele_counter<-0
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116 diviserror_counter<-0
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117 for(i in c(1:nrow(vcf_rtable))){
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118 vcf_line<-vcf_rtable[i,]
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119 #remove chrom or chr prefix from chromosome value
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120 if(grepl("chrom",vcf_line$CHROM,ignore.case=TRUE)==TRUE){
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121 vcf_line$CHROM<-gsub("chrom","",vcf_line$CHROM,ignore.case=TRUE)
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122 }else if(grepl("chr",vcf_line$CHROM,ignore.case=TRUE)==TRUE){
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123 vcf_line$CHROM<-gsub("chr","",vcf_line$CHROM,ignore.case=TRUE)
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124 }
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125 #PARSE INFO
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126 vcfinfo_split<-strsplit(vcf_line$INFO,split=";")
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127 vcfinfo_coln<-gsub("=.*","",unlist(vcfinfo_split))
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128 vcfinfo_cold<-gsub(".*=","",unlist(vcfinfo_split))
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129 vcfinfo_ldat<-data.frame(t(vcfinfo_cold),stringsAsFactors=FALSE)
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130 names(vcfinfo_ldat)<-vcfinfo_coln
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131
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132 #skip if commas in values to avoid returning errors
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133 if(grepl(",",vcfinfo_ldat$AO)==TRUE){
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134 multiallele_counter<-multiallele_counter+1
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135 next
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136 }
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137 #skip divide by zero errors (under "ratio" setting for ratio calculation)
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138 if(as.numeric(vcfinfo_ldat$AO)+as.numeric(vcfinfo_ldat$RO)=="0"){
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139 diviserror_counter<-diviserror_counter+1
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140 next
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141 }
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142
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143 #specific accounting for nonstandard categories
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144 #LOF columns only present for loss-of-function variants + assign NA values to all other variants
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145 if(("LOF" %in% names(vcfinfo_ldat))==TRUE){
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146 LOF<-vcfinfo_ldat$LOF
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147 vcfinfo_ldat<-vcfinfo_ldat[,!names(vcfinfo_ldat) %in% "LOF"]
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148 vcfinfo_ldat<-cbind(vcfinfo_ldat,LOF)
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149 }else{
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150 LOF<-"NA"
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151 vcfinfo_ldat<-cbind(vcfinfo_ldat,LOF)
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152 }
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153 #NMD columns only present for nonsense-mediated-decay variants + assign NA values to all other variants
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154 if(("NMD" %in% names(vcfinfo_ldat))==TRUE){
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155 NMD<-vcfinfo_ldat$NMD
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156 vcfinfo_ldat<-vcfinfo_ldat[,!names(vcfinfo_ldat) %in% "NMD"]
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157 vcfinfo_ldat<-cbind(vcfinfo_ldat,NMD)
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158 }else{
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159 NMD<-"NA"
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160 vcfinfo_ldat<-cbind(vcfinfo_ldat,NMD)
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161 }
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162 #PARSE ANNOTATION
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163 ann_rparsed<-unlist(strsplit(vcfinfo_ldat$ANN[1],split="\\|"))[1:20]
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164 ann_rparsed[ann_rparsed==""]<-"novalue"
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165 ann_parsed<-data.frame(t(ann_rparsed),stringsAsFactors=FALSE)
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166 names(ann_parsed)<-paste("ANN",c(1:dim(ann_parsed)[2]),sep="")
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167 #remove duplicate redundant INFO column (fully parsed)
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168 vcf_line<-vcf_line[,names(vcf_line)!="INFO"]
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169
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170 #dataset keeping parsed annotation (partial parsed-for relevant)
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171 vcfinfo_ldat<-vcfinfo_ldat[,names(vcfinfo_ldat)!="ANN"]
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172 #append copy of original data to parsed data
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173 vcfinfo_lldat<-cbind(vcf_line,vcfinfo_ldat,ann_parsed)
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174 vcfinfo_dat<-rbind(vcfinfo_dat,vcfinfo_lldat)
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175
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176 }
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177
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178 print(paste("rows with multiple alleles skipped: ",multiallele_counter,sep=""))
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179 # print(paste("rows with AO+RO=0 (not multiple alleles) skipped: ",diviserror_counter,sep=""))
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180
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181 #ENSURE CORRECT DATATYPES
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182 #convert dataframe columns of factor type to character type
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183 vcfinfo_dat<-data.frame(lapply(vcfinfo_dat,as.character),stringsAsFactors=FALSE)
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184 #convert to numeric if column is numeric and not string
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185 for(n in c(1:dim(vcfinfo_dat)[2])){
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186 #suppress warnings when columns with strings encountered (not converted)
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187 suppressWarnings(
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188 colnum_index<-!is.na(as.numeric(vcfinfo_dat[,n]))
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189 )
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190 if(all(colnum_index)==TRUE){
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191 vcfinfo_dat[,n]<-as.numeric(vcfinfo_dat[,n])
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192 }
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193 }
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194
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195
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196 ######################
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197 #RATIO CALCULATION
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198 #ratio calculation from AO and RO
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199 RATIO<-c(vcfinfo_dat$AO/(vcfinfo_dat$AO+vcfinfo_dat$RO))
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200 #add adj_AB for AB=0->AO=1 conversion
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201 adj_AB<-replace(vcfinfo_dat$AB,vcfinfo_dat$AB==0,1)
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202 vcfinfo_dat<-cbind(vcfinfo_dat,RATIO,adj_AB)
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203 vcfinfo_dat<-vcfinfo_dat[with(vcfinfo_dat,order(CHROM,POS)),]
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204
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205 vcfinfo_pdat<-vcfinfo_dat
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206 vcfinfo_dat<-subset(vcfinfo_dat,QUAL>=read_qual)
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207
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208 #CONSIDER ONLY SNP VARIANTS
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209 if(snp_only==TRUE){
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210 vcfinfo_dat<-subset(vcfinfo_dat,CIGAR=="1X")
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211 }
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212
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213 ######################
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214 #SUBTRACT VARIANTS FROM EXCLUSION LIST
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215 if(exclusion_list!="FALSE"){
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216 #keep copy of pre-subtraction data for later plotting
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217 vcfinfo_origdat<-vcfinfo_dat
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218
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219 #identifiers for exclusion list based on CHROM/POS/REF/ALT
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220 index1<-paste(vcfinfo_dat$CHROM,vcfinfo_dat$POS,sep="_")
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221 index2<-paste(vcfinfo_dat$CHROM,vcfinfo_dat$POS,vcfinfo_dat$REF,sep="_")
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222 index3<-paste(vcfinfo_dat$CHROM,vcfinfo_dat$POS,vcfinfo_dat$REF,vcfinfo_dat$ALT,sep="_")
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223 vcfinfo_dat<-cbind(vcfinfo_dat,index1,index2,index3)
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224
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225 print(paste("before subtraction: ",nrow(vcfinfo_dat),sep=""))
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226 #loop and subtract through exclusion lists (if multiple files)
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227 for(exclusion_ind in exclusion_list){
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228 exclin<-read.table(exclusion_ind,header=TRUE)
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229
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230 #THRESHOLD FILTER ON EXCLUSION LIST VARIANTS
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231 if(allele_ratio=="AB"){
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232 exclin<-subset(exclin,adj_AB>=excl_rthreshold_lower)
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233 exclin<-subset(exclin,adj_AB<=excl_rthreshold_upper)
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234 }
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235 if(allele_ratio=="ratio"){
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236 exclin<-subset(exclin,ratio>=excl_rthreshold_lower)
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237 exclin<-subset(exclin,ratio>=excl_rthreshold_upper)
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238 }
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239
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240 #identifiers for vcf data based on CHROM/POS/REF/ALT
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241 index1<-paste(exclin$CHROM,exclin$POS,sep="_")
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242 index2<-paste(exclin$CHROM,exclin$POS,exclin$REF,sep="_")
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243 index3<-paste(exclin$CHROM,exclin$POS,exclin$REF,exclin$ALT,sep="_")
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244 exclin<-cbind(exclin,index1,index2,index3)
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245 #exclude based on CHROM+POS+REF+ALT
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246 vcfinfo_dat<-subset(vcfinfo_dat,!(index3 %in% exclin$index3))
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247 }
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248 print(paste("after subtraction: ",nrow(vcfinfo_dat),sep=""))
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249 }
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250
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251
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252 ######################
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253 #WRITE TO OUTPUT
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254 #select relevant columns 2 variant type; 4 gene; !5 wormbase ID; 8 type change; 10 nucleotide change; 11 amino acid change; 16 warning message
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255 vcfinfo_simp<-subset(vcfinfo_dat,select=c("CHROM","POS","QUAL","DP","REF","ALT","AB","AO","RO","RATIO","adj_AB","ANN2","ANN4","ANN8","ANN10","ANN11","ANN16"))
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256 names(vcfinfo_simp)<-c("CHROM","POS","QUAL","DP","REF","ALT","AB","AO","RO","RATIO","adj_AB","VARTYPE","GENE","TYPE","NTCHANGE","PRCHANGE","WARNINGS")
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257 vcfsimp_dat<-vcfinfo_simp[with(vcfinfo_simp,order(CHROM,POS)),]
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258
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259 #write table with quality filtered variants for VDM plotting and relevant columns
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260 try(
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261 write.table(vcfsimp_dat,vcfoutput_filename,sep="\t",quote=FALSE,row.names=FALSE)
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262 ,silent=TRUE)
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263 # #write table with all unfiltered variants and all columns including parsed INFO
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264 # if(fastmode!="TRUE"){}
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265 # try(
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266 # write.table(vcfinfo_pdat,vcfparsed_filename,sep="\t",quote=FALSE,row.names=FALSE)
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267 # ,silent=TRUE)
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268 # }
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269
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270 ######################
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271 ###CHROMOSOME (INTERVAL) ARRANGEMENT
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272 #define chromosome and chromosome size in Mb
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273 if(interval_type == 'C.elegans'){
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274 chrom_n<-c('I','II','III','IV','V','X')
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275 chrom_mb<-c(16,16,14,18,21,18)
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276 interval_frame<-data.frame(chrom_n,chrom_mb)
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277 } else if(interval_type == 'Zebrafish'){
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278 chrom_n<-c('1','2','3','4','5','6','7','8','9','10','11','12','13','14','15','16','17','18','19','20','21','22','23','24','25')
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279 chrom_mb<-c(61,61,64,63,76,60,78,57,59,47,47,51,55,54,48,59,54,50,51,56,45,43,47,44,39)
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280 interval_frame<-data.frame(chrom_n,chrom_mb)
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281 } else if(interval_type == 'Brachypodium'){
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282 chrom_n<-c('1','2','3','4','5')
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283 chrom_mb<-c(75,60,60,50,30)
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284 interval_frame<-data.frame(chrom_n,chrom_mb)
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285 } else if(interval_type == 'Arabidopsis'){
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286 chrom_n<-c('1','2','3','4','5')
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287 chrom_mb<-c(31, 20,24,19,27 )
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288 interval_frame<-data.frame(chrom_n,chrom_mb)
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289 } else{
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290 #user interval file- no headers, with chromosome in column 1 (format CHR# or CHROM#) and size in Mb (rounded up) in column 2
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291 user_interval_type<-read.table(interval_type)
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292 if(grepl("chrom",user_interval_type[1,1],ignore.case=TRUE)==TRUE){
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293 user_interval_type[,1]<-gsub("chrom","",user_interval_type[,1],ignore.case=TRUE)
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294 }else if(grep("chr",user_interval_type[1,1],ignore.case=TRUE)==TRUE){
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295 user_interval_type[,1]<-gsub("chr","",user_interval_type[,1],ignore.case=TRUE)
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296 }
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297 chrom_n<-user_interval_type[,1]
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298 chrom_mb<-user_interval_type[,2]
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299 interval_frame<-data.frame(chrom_n,chrom_mb)
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300 }
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301 names(interval_frame)<-c("CHROM","INTERVAL")
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302
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303
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304 ######################
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305 ###PLOTTING
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306 #VDM SCATTER PLOT
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307 #save to pdf
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308 pdf(file=pdf_filename,width=9,height=8)
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309 #par(mfrow=c(2,3))
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310 for(chromind in interval_frame$CHROM){
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311 #subset by data by chromosome for plotting
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312 intervalind<-interval_frame$INTERVAL[interval_frame$CHROM==chromind]
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313 chr_dat<-subset(vcfsimp_dat,CHROM==chromind,silent=TRUE)
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314
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315 #same subsetting by chromosome for pre-subtraction data
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316 if(exclusion_list!="FALSE"){
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317 chr_origdat<-subset(vcfinfo_origdat,CHROM==chromind,silent=TRUE)
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318 }
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319
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320 #define x-axis upper limit
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321 if(xaxis_standard==TRUE){
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322 #for standardized x-axis (max x-axis chromosome length)
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323 scupper_xaxis<-max(interval_frame$INTERVAL)
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324 scupper_xval<-scupper_xaxis*interval_unit
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325 } else if(xaxis_standard==FALSE){
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326 scupper_xaxis<-intervalind
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327 scupper_xval<-intervalind*interval_unit
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328 }
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329
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330 if(allele_ratio=="AB"){
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331 plot(chr_dat$POS,chr_dat$adj_AB,cex=0.60,xlim=c(0,scupper_xval),ylim=c(0,sp_yaxis),main=paste("Chr",chromind," Variant Discovery Mapping",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Ratio of Variant Reads/Total Reads [AB]',pch=10, col=plot_color,xaxt='n')
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332 try(lines(loess.smooth(chr_dat$POS,chr_dat$adj_AB,span=loess_span),lwd=5,col=loess_color))
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333 #plot loess curve for data without subtraction of exclusion variants
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334 if(exclusion_list!="FALSE"){
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335 try(lines(loess.smooth(chr_origdat$POS,chr_origdat$adj_AB,span=loess_span),lty="longdash",lwd=4,col=excl_loess_color))
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336 }
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337
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338 axis(1,at=seq(0,scupper_xval,by=interval_unit),labels=c(0:scupper_xaxis))
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339 abline(h=seq(0,sp_yaxis,by=0.1),v=c(1:scupper_xaxis)*interval_unit,col="gray")
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340 } else if(allele_ratio=="ratio"){
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341 plot(chr_dat$POS,chr_dat$RATIO,cex=0.60,xlim=c(0,scupper_xval),ylim=c(0,sp_yaxis),main=paste("Chr",chromind," Variant Discovery Mapping",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Ratio of Variant Reads/Total Reads [ratio]',pch=10, col=plot_color,xaxt='n')
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342 try(lines(loess.smooth(chr_dat$POS,chr_dat$RATIO,span=loess_span),lwd=5,col=loess_color))
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343 #plot loess curve for data without subtraction of exclusion variants
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344 if(exclusion_list!="FALSE"){
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345 try(lines(loess.smooth(chr_origdat$POS,chr_origdat$adj_AB,span=loess_span),lty="longdash",lwd=4,col=excl_loess_color))
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346 }
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347 axis(1,at=seq(0,scupper_xval,by=interval_unit),labels=c(0:scupper_xaxis))
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348 abline(h=seq(0,sp_yaxis,by=0.1),v=c(1:scupper_xaxis)*interval_unit,col="gray")
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349 }
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350 }
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351
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352
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353 ######################
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354 #graph barplots
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355 location_index<-NULL
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356 meanSNP_dat<-NULL
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357 # prepare table of counts and calculations
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358 for(chromind in interval_frame$CHROM){
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359 #for standardized x-axis
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360 if(xaxis_standard==TRUE){
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361 intervalind<-max(interval_frame$INTERVAL)*1000000/bin_size
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362 } else if(xaxis_standard==FALSE){
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363 intervalind<-interval_frame$INTERVAL[interval_frame$CHROM==chromind]*1000000/bin_size
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364 }
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365 #start intervals with **1 and end with **0
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366 interval_begin<-c(((0:(intervalind-1))*bin_size)+1)
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367 interval_end<-c((1:intervalind)*bin_size)
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368
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369 #define x-axis upper limit
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370 if(xaxis_standard==TRUE){
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371 upper_xaxis<-max(interval_frame$INTERVAL)
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372 } else if(xaxis_standard==FALSE){
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373 upper_xaxis<-interval_frame$INTERVAL[interval_frame$CHROM==chromind]
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374 }
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375 #prepare columns
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376 snp_counter<-0
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377 purealt_counter<-0
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378 pureref_counter<-0
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379 het_counter<-0
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380 chr_mean<-0
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381 normed_freq<-0
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382 ratio<-0
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383
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384 interval_index<-data.frame(chromind,interval_begin,interval_end,snp_counter,purealt_counter,pureref_counter,het_counter,chr_mean,normed_freq)
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385 chr_dat<-subset(vcfinfo_dat,CHROM==chromind)
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386 #ratio calculation
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387 ratio<-chr_dat$AO/(chr_dat$AO+chr_dat$RO)
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388
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389 #if counter based on adj_AB or ratio
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390 if(allele_ratio=="ratio"){
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391 chr_purealtdat<-subset(chr_dat,ratio>=threshold_upper)
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392 chr_purerefdat<-subset(chr_dat,ratio<=threshold_lower)
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393 chr_hetdat<-subset(chr_dat,ratio>threshold_lower & ratio<threshold_upper)
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394 } else if(allele_ratio=="AB"){
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395 chr_purealtdat<-subset(chr_dat,adj_AB>=threshold_upper)
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396 chr_purerefdat<-subset(chr_dat,adj_AB<=threshold_lower)
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397 chr_hetdat<-subset(chr_dat,adj_AB>threshold_lower & adj_AB<threshold_upper)
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398 }
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399 #if chromosome with data, count number of snps within each bin (positions rounded up to nearest bin), else skip to next chromosome
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400 if(dim(chr_dat)[1]>0){
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401 for(i in 1:dim(chr_dat)[1]){
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402 chr_datind<-chr_dat[i,]
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403 #round up to nearest bin-size interval
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404 chr_datind_upper<-ceiling(chr_datind$POS/bin_size)*bin_size
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405 interval_coln<-NULL;interval_rown<-NULL
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406 #identify row and and counter column to increment
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407 interval_coln<-which(names(interval_index)=="snp_counter")
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408 interval_rown<-match(chr_datind_upper,interval_index$interval_end)
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409 interval_index[interval_rown,interval_coln]<-c(interval_index$snp_counter[interval_rown]+1)
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410 }
|
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411 }else{
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412 next
|
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413 }
|
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414 #if chromosome with pure AO, count number of snps with each bin (positions rounded up to nearest bin)
|
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415 if(dim(chr_purealtdat)[1]>0){
|
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416 for(i in 1:dim(chr_purealtdat)[1]){
|
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417 chr_purealtind<-chr_purealtdat[i,]
|
|
418 chr_purealtind_upper<-ceiling(chr_purealtind$POS/bin_size)*bin_size
|
|
419 interval_coln<-NULL;interval_rown<-NULL
|
|
420 interval_coln<-which(names(interval_index)=="purealt_counter")
|
|
421 interval_rown<-match(chr_purealtind_upper,interval_index$interval_end)
|
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422 interval_index[interval_rown,interval_coln]<-c(interval_index$purealt_counter[interval_rown]+1)
|
|
423 }
|
|
424 }
|
|
425 #if chromosome with pure RO, count number of snps with each bin (positions rounded up to nearest bin)
|
|
426 if(dim(chr_purerefdat)[1]>0){
|
|
427 for(i in 1:dim(chr_purerefdat)[1]){
|
|
428 chr_purerefind<-chr_purerefdat[i,]
|
|
429 chr_purerefind_upper<-ceiling(chr_purerefind$POS/bin_size)*bin_size
|
|
430 interval_coln<-NULL;interval_rown<-NULL
|
|
431 interval_coln<-which(names(interval_index)=="pureref_counter")
|
|
432 interval_rown<-match(chr_purerefind_upper,interval_index$interval_end)
|
|
433 interval_index[interval_rown,interval_coln]<-c(interval_index$pureref_counter[interval_rown]+1)
|
|
434 }
|
|
435 }
|
|
436 #if chromosome with hets, count number of snps with each bin (positions rounded up to nearest bin)
|
|
437 if(dim(chr_hetdat)[1]>0){
|
|
438 for(i in 1:dim(chr_hetdat)[1]){
|
|
439 chr_hetind<-chr_hetdat[i,]
|
|
440 chr_hetind_upper<-ceiling(chr_hetind$POS/bin_size)*bin_size
|
|
441 interval_coln<-NULL;interval_rown<-NULL
|
|
442 interval_coln<-which(names(interval_index)=="het_counter")
|
|
443 interval_rown<-match(chr_hetind_upper,interval_index$interval_end)
|
|
444 interval_index[interval_rown,interval_coln]<-c(interval_index$het_counter[interval_rown]+1)
|
|
445 }
|
|
446 }
|
|
447 #irrespective of standardized x-axis, mean should be calculated from actual interval range of chromosome
|
|
448 chr_mean<-sum(interval_index$purealt_counter)/(interval_frame$INTERVAL[interval_frame$CHROM==chromind]*1000000/bin_size)
|
|
449 interval_index$chr_mean<-chr_mean
|
|
450 meanSNP_dat<-rbind(meanSNP_dat,data.frame(chromind,chr_mean))
|
|
451 #normalization treatment for if SNPs are AO=0, AO=total SNPs, or AO and RO in bin
|
|
452 for(i in 1:dim(interval_index)[1]){
|
|
453 chr_intind<-interval_index[i,]
|
|
454 #hom definition based on specified upper and lower thresholds
|
|
455 if(chr_intind$purealt_counter<=threshold_lower){
|
|
456 interval_coln<-NULL
|
|
457 interval_coln<-which(names(interval_index)=="normed_freq")
|
|
458 interval_index[i,interval_coln]=0
|
|
459 } else if (chr_intind$purealt_counter==chr_intind$snp_counter){
|
|
460 interval_coln<-NULL
|
|
461 interval_coln<-which(names(interval_index)=="normed_freq")
|
|
462 interval_index[i,interval_coln]=(chr_intind$purealt_counter)^2/chr_mean
|
|
463 } else {
|
|
464 interval_coln<-NULL
|
|
465 interval_coln<-which(names(interval_index)=="normed_freq")
|
|
466 interval_index[i,interval_coln]=chr_mean*(chr_intind$purealt_counter)^2/(chr_intind$snp_counter-chr_intind$purealt_counter)
|
|
467 }
|
|
468 }
|
|
469 location_index<-rbind(location_index,interval_index)
|
|
470 }
|
|
471
|
|
472 for(chromind in interval_frame$CHROM){
|
|
473 interval_index<-location_index[location_index$chromind==chromind,]
|
|
474 #assign 0 values to avoid empty datatable error
|
|
475 if(dim(interval_index)[1]==0){
|
|
476 interval_index[1,]<-rep(0,dim(interval_index)[2])
|
|
477 }
|
|
478 #set up x_axis
|
|
479 if(xaxis_standard==TRUE){
|
|
480 #for standardized x-axis (max x-axis chromosome length)
|
|
481 bpupper_xaxis<-max(interval_frame$INTERVAL)
|
|
482 bpupper_xval<-bpupper_xaxis*interval_unit
|
|
483 }else if(xaxis_standard==FALSE){
|
|
484 bpupper_xaxis<-intervalind
|
|
485 bpupper_xval<-intervalind*interval_unit
|
|
486 }
|
|
487 #set up y_axis range for barplots
|
|
488 if(bfreq_norm==TRUE){
|
|
489 bp_yaxis<-5*ceiling(max(location_index$normed_freq)/5)
|
|
490 #assign non-0 value to yaxis to avoid error
|
|
491 if(bp_yaxis==0){
|
|
492 bp_yaxis<-10
|
|
493 }
|
|
494 if(xaxis_standard==TRUE){
|
|
495 bplot<-barplot(interval_index$normed_freq,space=0,ylim=c(0,bp_yaxis),main=paste("Chr",chromind," Variant Only",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Normalised Frequency')
|
|
496 }else if(xaxis_standard==FALSE){
|
|
497 bplot<-barplot(interval_index$normed_freq,space=0,xlim=c(0,bpupper_xaxis),ylim=c(0,bp_yaxis),main=paste("Chr",chromind," Variant Only",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Normalised Frequency')
|
|
498 }
|
|
499
|
|
500 }else if(bfreq_norm==FALSE){
|
|
501 bp_yaxis<-5*ceiling(max(location_index$purealt_counter)/5)
|
|
502 #assign non-0 value to yaxis to avoid error
|
|
503 if(bp_yaxis==0){
|
|
504 bp_yaxis<-10
|
|
505 }
|
|
506 if(xaxis_standard==TRUE){
|
|
507 bplot<-barplot(interval_index$purealt_counter,space=0,ylim=c(0,bp_yaxis),main=paste("Chr",chromind," Variant Only",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Frequency')
|
|
508 }else if(xaxis_standard==FALSE){
|
|
509 bplot<-barplot(interval_index$purealt_counter,space=0,xlim=c(0,bpupper_xaxis),ylim=c(0,bp_yaxis),main=paste("Chr",chromind," Variant Only",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Frequency')
|
|
510 }
|
|
511 }
|
|
512 bp_xaxis1<-as.numeric(bplot)
|
|
513 bp_xaxis2<-c(bp_xaxis1,tail(bp_xaxis1,1)+bp_xaxis1[2]-bp_xaxis1[1])
|
|
514 bp_xaxis<-bp_xaxis2-bp_xaxis1[1]
|
|
515
|
|
516 axis(1,at=bp_xaxis,labels=seq(0,bpupper_xaxis,by=c(bin_size/1000000)))
|
|
517 }
|
|
518 dev.off()
|
|
519 }
|
|
520
|
|
521 myfunction(inf=options$inf,itype=options$itype,qual=options$qual,allr=options$allr,snp=options$snp,lsp=options$lsp,pcol=options$pcol,lcol=options$lcol,
|
|
522 xstand=options$xstand,bsize=options$bsize,bnorm=options$bnorm,freqthr=options$freqthr,exclf=options$exclf,exclcol=options$exclcol,outn=options$outn,pdfn=options$pdfn)
|
|
523
|