changeset 1:48e2fe881400 draft

Deleted selected files

--- a/my_VDM_tool.r	Thu Nov 05 13:48:08 2020 +0000
+++ /dev/null	Thu Jan 01 00:00:00 1970 +0000
@@ -1,604 +0,0 @@
-#!/usr/bin/env Rscript
-
-# rm(list=ls())
-
-library("getopt")
-
-#input from trailing line arguments
-args <- commandArgs(trailingOnly = TRUE)
-#read the options from input commandArgs
-option_specification = matrix(c(
-  'inf','i01',1,'character',
-  'itype','i02',2,'character',
-  'thrup','i03',2,'double',
-  'thrlow','i04',2,'double',
-  'allr','i05',2,'character',
-  'complex','i06',2,'logical',
-  'lsp','i07',2,'double',
-  'pcol','i08',2,'character',
-  'lcol','i09',2,'character',
-  
-  'xstand','i10',2,'logical',
-  'bsize','i11',2,'integer',
-  'bnorm','i12',2,'logical',
-  'exclthr','i13',2,'double',
-  'exclcol','i14',2,'character',
-  
-  'parn','o1',2,'character',
-  'outn','o2',2,'character',
-  'pdfn','o3',2,'character'
-), byrow=TRUE, ncol=4)
-
-#parse options
-options = getopt(option_specification)
-
-# #FOR DEBUGGING
-# options<-NULL
-# ###INPUT FILE
-# setwd("D:/Dropbox/_galaxy/")
-# options$inf<-"nor22.vcf"
-# ###BASE OPTIONS
-# #interval type
-# options$itype<-"C.elegans"
-# #user interval type
-# options$userif<-NULL
-# #quality filter
-# options$qual<-200
-# #for scaling 0-1 = upper threshold for what is considered homozygous
-# options$thrup<-1
-# #for scaling 0-1 = lower threshold for what is considered homozygous
-# options$thrlow<-0
-# #type of allelic ratio (AB/ratio)+---------------------------------------------------------
-# options$allr<-"AB"
-# #include complex variants
-# options$complex<-FALSE
-# ###ADDITIONAL VARIANT EXCLUSION OPTIONS
-# #files with variants to exclude
-# options$exclf<-NULL
-# options$exclf<-c("nor22chunk1.txt","nor22chunk5.txt")
-# #for variants to exclude, bottom threshold for which to be used, i.e. 0=ALL, 1=HOM only, 0.8=near HOM)
-# options$exclthr<-0
-# #additional colour option for pre-subtraction line
-# options$exclcol<-"green"
-# ###PLOT OPTIONS
-# #loess span
-# options$lsp<-0.4
-# #point colour
-# options$pcol<-"black"
-# #loess plot colour
-# options$lcol<-"red"
-# #standardize x-axis interval (e.g. 1Mb interval)
-# options$xstand<-TRUE
-# #bin size for barplot
-# options$bsize<-1000000
-# #normalization for barplot
-# options$bnorm<-FALSE
-# ###OUTPUT OPTIONS
-# #custom files names (may not work for Galaxy)
-# options$outn<-paste(gsub("vcf","",options$inf),"_output_q",options$qual,"-",paste(options$exclf,sep="",collapse=""),".txt",sep="")
-# options$parn<-paste(gsub("vcf","",options$inf),"_parsed_q",options$qual,"-",paste(options$exclf,sep="",collapse=""),".txt",sep="")
-# options$pdfn<-paste(gsub("vcf","",options$inf),"_plot_q",options$qual,"-",paste(options$exclf,sep="",collapse=""),".pdf",sep="")
-# #fixed file names (will work in Galaxy)
-# # options$outn<-"vcf_output.txt"
-# # options$parn<-"vcf_parsedinput.txt"
-# # options$pdfn<-"vdm_mapping_plot.pdf"
-
-
-'inf','i01',1,'character',
-'itype','i02',2,'character',
-'qual','i03',2,'double',
-'thrup','i04',2,'double',
-'thrlow','i05',2,'double',
-'allr','i06',2,'character',
-'complex','i07',2,'logical',
-'lsp','i08',2,'double',
-'pcol','i09',2,'character',
-'lcol','i10',2,'character',
-
-'xstand','i11',2,'logical',
-'bsize','i12',2,'integer',
-'bnorm','i13',2,'logical',
-'exclthr','i14',2,'double',
-'exclcol','i15',2,'character',
-
-'parn','o1',2,'character',
-'outn','o2',2,'character',
-'pdfn','o3',2,'character'
-
-
-myfunction<-function(inf,itype,thrup,thrlow,allr,complex,lsp,pcol,lcol,xstand,bsize,bnorm,exclthr,exclcol,parn,outn,pdfn){
-  
-  #PARAMETERS
-  # filename<-options$inf
-  # interval_type<-options$itype
-  # user_interval_file<-options$userif
-  # read_qual<-options$qual
-  # threshold_upper<-options$thrup
-  # threshold_lower<-options$thrlow
-  # allele_ratio<-options$allr
-  # incl_complex<-options$complex
-  # loess_span<-options$lsp
-  # plot_color<-options$pcol
-  # loess_color<-options$lcol
-  # #transparency for selected colour (to see plot points underneath)
-  # xaxis_standard<-options$xstand
-  # bin_size<-options$bsize
-  # bfreq_norm<-options$bnorm
-  # exclusion_list<-options$exclf
-  # excl_threshold<-options$exclthr
-  # excl_loess_color<-options$exclcol
-  # vcfoutput_filename<-options$outn
-  # vcfparsed_filename<-options$parn
-  # pdf_filename<-options$pdfn
-
-  # plot_color<-rgb(c(col2rgb(plot_color)[1]),c(col2rgb(plot_color)[2]),c(col2rgb(plot)[3]),max=255,alpha=150)
-  # loess_color<-rgb(c(col2rgb(loess_color)[1]),c(col2rgb(loess_color)[2]),c(col2rgb(loess_color)[3]),max=255,alpha=150)
-  # 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)
-  
-  inf,itype,thrup,thrlow,allr,complex,lsp,pcol,lcol,xstand,bsize,bnorm,exclthr,exclcol,parn,outn,pdfn
-  
-  filename<-inf
-  interval_type<-itype
-  # user_interval_file<-userif
-  read_qual<-as.numeric(qual)
-  threshold_upper<-as.numeric(thrup)
-  threshold_lower<-as.numeric(thrlow)
-  allele_ratio<-allr
-  incl_complex<-complex
-  loess_span<-as.numeric(lsp)
-  plot_color<-pcol
-  loess_color<-lcol
-  xaxis_standard<-xstand
-  bin_size<-as.numeric(bsize)
-  bfreq_norm<-bnorm
-  # exclusion_list<-exclf
-  excl_threshold<-as.numeric(exclthr)
-  excl_loess_color<-exclcol
-  vcfoutput_filename<-outn
-  vcfparsed_filename<-parn
-  pdf_filename<-pdfn
-
-  #transparency for selected colour (to see plot points underneath)
-  plot_color<-rgb(c(col2rgb(plot_color)[1]),c(col2rgb(plot_color)[2]),c(col2rgb(plot)[3]),max=255,alpha=150)
-  loess_color<-rgb(c(col2rgb(loess_color)[1]),c(col2rgb(loess_color)[2]),c(col2rgb(loess_color)[3]),max=255,alpha=150)
-  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)
-  
-  ###FIXED PARAMETERS
-  #linkage scatter plot yaxis max value=1
-  sp_yaxis<-1 
-  #chromosome intervals in Mb rather than custom
-  interval_unit<-1000000 
-
-
-######################
-###READ IN VCF FILE
-#extract column names
-vcf_readin<-readLines(filename)   
-#find header line, i.e. last line to begin with #
-for(l in 1:length(vcf_readin)){
-  vcf_readinl<-vcf_readin[l]
-  if(substr(vcf_readinl,1,1)=="#"){next}
-  else if(substr(vcf_readinl,1,1)!="#"){rowline<-l-1;break}
-}
-vcf_header<-vcf_readin[rowline]
-#e.g. CHROM\tPOS\tID\tREF\tALT\tQUAL\tFILTER\tINFO\tFORMAT\trgSM"
-vcf_header<-gsub("#","",vcf_header)
-vcf_colnames<-unlist(strsplit(vcf_header,"\t"))
-
-#extract data (hashed vcf header skipped with read.table)
-vcf_rtable<-read.table(filename,sep="\t",stringsAsFactors=FALSE)
-names(vcf_rtable)<-vcf_colnames
-
-######################
-###PREPARE DATA
-
-vcfinfo_dat<-NULL
-vcfinfo_pdat<-NULL
-multiallele_counter<-0
-diviserror_counter<-0
-for(i in c(1:nrow(vcf_rtable))){
-  vcf_line<-vcf_rtable[i,]
-  #to speed up runtime- quality filter here
-  if(vcf_line$QUAL>=read_qual){
-    #remove chrom or chr prefix from chromosome value
-    if(grepl("chrom",vcf_line$CHROM,ignore.case=TRUE)==TRUE){
-      vcf_line$CHROM<-gsub("chrom","",vcf_line$CHROM,ignore.case=TRUE)
-    }else if(grepl("chr",vcf_line$CHROM,ignore.case=TRUE)==TRUE){
-      vcf_line$CHROM<-gsub("chr","",vcf_line$CHROM,ignore.case=TRUE)
-    }
-#PARSE INFO
-    vcfinfo_split<-strsplit(vcf_line$INFO,split=";")	
-    vcfinfo_coln<-gsub("=.*","",unlist(vcfinfo_split))
-    vcfinfo_cold<-gsub(".*=","",unlist(vcfinfo_split))
-    vcfinfo_ldat<-data.frame(t(vcfinfo_cold),stringsAsFactors=FALSE)
-    names(vcfinfo_ldat)<-vcfinfo_coln
-    
-    #skip if commas in values to avoid returning errors
-    if(grepl(",",vcfinfo_ldat$AO)==TRUE){
-      multiallele_counter<-multiallele_counter+1
-      next
-    }
-    #skip divide by zero errors (under "ratio" setting for ratio calculation)
-    if(as.numeric(vcfinfo_ldat$AO)+as.numeric(vcfinfo_ldat$RO)=="0"){
-      diviserror_counter<-diviserror_counter+1
-      next
-    }		
-    
-    #specific accounting for nonstandard categories    
-    #LOF columns only present for loss-of-function variants + assign NA values to all other variants
-    if(("LOF" %in% names(vcfinfo_ldat))==TRUE){
-      LOF<-vcfinfo_ldat$LOF
-      vcfinfo_ldat<-vcfinfo_ldat[,!names(vcfinfo_ldat) %in% "LOF"]
-      vcfinfo_ldat<-cbind(vcfinfo_ldat,LOF)
-    }else{
-      LOF<-"NA"
-      vcfinfo_ldat<-cbind(vcfinfo_ldat,LOF)
-    }
-    #NMD columns only present for nonsense-mediated-decay variants + assign NA values to all other variants
-    if(("NMD" %in% names(vcfinfo_ldat))==TRUE){
-      NMD<-vcfinfo_ldat$NMD
-      vcfinfo_ldat<-vcfinfo_ldat[,!names(vcfinfo_ldat) %in% "NMD"]
-      vcfinfo_ldat<-cbind(vcfinfo_ldat,NMD)
-    }else{
-      NMD<-"NA"
-      vcfinfo_ldat<-cbind(vcfinfo_ldat,NMD)
-    }
-    
-    #general accounting for nonstandard categories
-  
-
-#PARSE ANNOTATION
-    ann_rparsed<-unlist(strsplit(vcfinfo_ldat$ANN[1],split="\\|"))[1:20]
-    ann_rparsed[ann_rparsed==""]<-"novalue"
-    ann_parsed<-data.frame(t(ann_rparsed),stringsAsFactors=FALSE)
-    names(ann_parsed)<-paste("ANN",c(1:dim(ann_parsed)[2]),sep="")
-    #remove duplicate redundant INFO column (fully parsed)
-    vcf_line<-vcf_line[,names(vcf_line)!="INFO"]
-    
-    #dataset keeping unparsed annotation (full)
-    vcfinfo_pldat<-cbind(vcf_line,vcfinfo_ldat)
-    vcfinfo_pdat<-rbind(vcfinfo_pdat,vcfinfo_pldat)
-    
-    #dataset keeping parsed annotation (partial parsed-for relevant)
-    vcfinfo_ldat<-vcfinfo_ldat[,names(vcfinfo_ldat)!="ANN"]
-    #append copy of original data to parsed data
-    vcfinfo_lldat<-cbind(vcf_line,vcfinfo_ldat,ann_parsed)
-    vcfinfo_dat<-rbind(vcfinfo_dat,vcfinfo_lldat)
-  }
-}
-print(paste("rows with multiple alleles skipped: ",multiallele_counter,sep=""))
-# print(paste("rows with AO+RO=0 (not multiple alleles) skipped: ",diviserror_counter,sep=""))
-
-#ENSURE CORRECT DATATYPES
-#convert dataframe columns of factor type to character type
-vcfinfo_dat<-data.frame(lapply(vcfinfo_dat,as.character),stringsAsFactors=FALSE)
-#convert to numeric if column is numeric and not string
-for(n in c(1:dim(vcfinfo_dat)[2])){
-  #suppress warnings when columns with strings encountered (not converted)
-  suppressWarnings(
-    colnum_index<-!is.na(as.numeric(vcfinfo_dat[,n]))
-  )
-  if(all(colnum_index)==TRUE){
-    vcfinfo_dat[,n]<-as.numeric(vcfinfo_dat[,n])
-  }
-}
-
-######################
-#RATIO CALCULATION
-#ratio calculation from AO and RO
-RATIO<-c(vcfinfo_dat$AO/(vcfinfo_dat$AO+vcfinfo_dat$RO))
-#add adj_AB for AB=0->AO=1 conversion
-adj_AB<-replace(vcfinfo_dat$AB,vcfinfo_dat$AB==0,1)
-vcfinfo_dat<-cbind(vcfinfo_dat,RATIO,adj_AB)
-vcfinfo_dat<-vcfinfo_dat[with(vcfinfo_dat,order(CHROM,POS)),]
-
-#EXCLUDE COMPLEX VARIANTS
-if(incl_complex==FALSE){
-  vcfinfo_dat<-subset(vcfinfo_dat,CIGAR=="1X")
-}
-
-######################
-#SUBTRACT VARIANTS FROM EXCLUSION LIST
-if(length(exclusion_list)>0){
-  #keep copy of pre-subtraction data for later plotting
-  vcfinfo_origdat<-vcfinfo_dat
-  
-  #identifiers for exclusion list based on CHROM/POS/REF/ALT
-  index1<-paste(vcfinfo_dat$CHROM,vcfinfo_dat$POS,sep="_")
-  index2<-paste(vcfinfo_dat$CHROM,vcfinfo_dat$POS,vcfinfo_dat$REF,sep="_")
-  index3<-paste(vcfinfo_dat$CHROM,vcfinfo_dat$POS,vcfinfo_dat$REF,vcfinfo_dat$ALT,sep="_")
-  vcfinfo_dat<-cbind(vcfinfo_dat,index1,index2,index3)
-  
-  print(paste("before: ",nrow(vcfinfo_dat),sep=""))
-  #loop and subtract through exclusion lists (if multiple files)
-  for(exclusion_ind in exclusion_list){
-    exclin<-read.table(exclusion_ind,header=TRUE)
-  
-  #THRESHOLD FILTER ON EXCLUSION LIST VARIANTS  
-    if(allele_ratio=="AB"){
-      exclin<-subset(exclin,adj_AB>=excl_threshold)
-    }
-    if(allele_ratio=="ratio"){
-      exclin<-subset(exclin,ratio>=excl_threshold)
-    }
-  
-    #identifiers for vcf data based on CHROM/POS/REF/ALT  
-    index1<-paste(exclin$CHROM,exclin$POS,sep="_")
-    index2<-paste(exclin$CHROM,exclin$POS,exclin$REF,sep="_")
-    index3<-paste(exclin$CHROM,exclin$POS,exclin$REF,exclin$ALT,sep="_")
-    exclin<-cbind(exclin,index1,index2,index3)
-    #exclude based on CHROM+POS+REF+ALT
-    vcfinfo_dat<-subset(vcfinfo_dat,!(index3 %in% exclin$index3))
-  }
-  print(paste("after: ",nrow(vcfinfo_dat),sep=""))
-}
-
-######################
-#WRITE TO OUTPUT
-#select relevant columns 2 variant type; 4 gene; !5 wormbase ID; 8 type change; 10 nucleotide change; 11 amino acid change; 16 warning message
-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"))
-names(vcfinfo_simp)<-c("CHROM","POS","QUAL","DP","REF","ALT","AB","AO","RO","RATIO","adj_AB","VARTYPE","GENE","TYPE","NTCHANGE","PRCHANGE","WARNINGS")
-vcfsimp_dat<-vcfinfo_simp[with(vcfinfo_simp,order(CHROM,POS)),]
-
-#write table with quality filtered variants for VDM plotting and relevant columns
-try(
-  write.table(vcfsimp_dat,vcfoutput_filename,sep="\t",quote=FALSE,row.names=FALSE)
-  ,silent=TRUE)
-#write table with all unfiltered variants and all columns including parsed INFO
-try(
-  write.table(vcfinfo_pdat,vcfparsed_filename,sep="\t",quote=FALSE,row.names=FALSE)
-  ,silent=TRUE)
-
-
-######################
-###CHROMOSOME (INTERVAL) ARRANGEMENT
-#define chromosome and chromosome size in Mb
-if(interval_type == 'C.elegans'){
-  chrom_n<-c('I','II','III','IV','V','X')
-  chrom_mb<-c(16,16,14,18,21,18)
-  interval_frame<-data.frame(chrom_n,chrom_mb)
-} else if(interval_type == 'Zebrafish'){
-  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')
-  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)
-  interval_frame<-data.frame(chrom_n,chrom_mb)
-} else if(interval_type == 'Brachypodium'){
-  chrom_n<-c('1','2','3','4','5')
-  chrom_mb<-c(75,60,60,50,30)
-  interval_frame<-data.frame(chrom_n,chrom_mb)
-} else if(interval_type == 'Arabidopsis'){
-  chrom_n<-c('1','2','3','4','5')
-  chrom_mb<-c(31, 20,24,19,27 )
-  interval_frame<-data.frame(chrom_n,chrom_mb)
-} else{
-  #user interval file- no headers, with chromosome in column 1 (format CHR# or CHROM#) and size in Mb (rounded up) in column 2
-  user_interval_type<-read.table(user_interval_file)
-  if(grepl("chrom",user_interval_type[1,1],ignore.case=TRUE)==TRUE){
-    user_interval_type[,1]<-gsub("chrom","",user_interval_type[,1],ignore.case=TRUE)
-  }else if(grep("chr",user_interval_type[1,1],ignore.case=TRUE)==TRUE){
-    user_interval_type[,1]<-gsub("chr","",user_interval_type[,1],ignore.case=TRUE)
-  }
-  chrom_n<-user_interval_type[,1]
-  chrom_mb<-user_interval_type[,2]
-  interval_frame<-data.frame(chrom_n,chrom_mb)
-}
-names(interval_frame)<-c("CHROM","INTERVAL")
-
-
-######################
-###PLOTTING
-#VDM SCATTER PLOT
-#save to pdf
-pdf(file=pdf_filename,width=9,height=8)
-#par(mfrow=c(2,3))
-for(chromind in interval_frame$CHROM){
-  #subset by data by chromosome for plotting
-  intervalind<-interval_frame$INTERVAL[interval_frame$CHROM==chromind]
-  chr_dat<-subset(vcfsimp_dat,CHROM==chromind,silent=TRUE)
-  
-  #same subsetting by chromosome for pre-subtraction data
-  if(length(exclusion_list)>0){
-    chr_origdat<-subset(vcfinfo_origdat,CHROM==chromind,silent=TRUE)
-  }
-  
-  #define x-axis upper limit
-  if(xaxis_standard==TRUE){
-    #for standardized x-axis (max x-axis chromosome length)
-    scupper_xaxis<-max(interval_frame$INTERVAL)
-    scupper_xval<-scupper_xaxis*interval_unit
-  } else if(xaxis_standard==FALSE){
-    scupper_xaxis<-intervalind
-    scupper_xval<-intervalind*interval_unit
-  }
-  
-  if(allele_ratio=="AB"){
-    plot(chr_dat$POS,chr_dat$adj_AB,cex=0.60,xlim=c(0,scupper_xval),ylim=c(0,sp_yaxis),main=paste("Chr",chromind," Variant Discovery Mapping",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Ratio of Variant Reads/Total Reads [AB]',pch=10, col=plot_color,xaxt='n')				
-    try(lines(loess.smooth(chr_dat$POS,chr_dat$adj_AB,span=loess_span),lwd=5,col=loess_color))
-    #plot loess curve for data without subtraction of exclusion variants
-    if(length(exclusion_list)>0){
-      try(lines(loess.smooth(chr_origdat$POS,chr_origdat$adj_AB,span=loess_span),lwd=5,col=excl_loess_color))
-    }
-    
-    axis(1,at=seq(0,scupper_xval,by=interval_unit),labels=c(0:scupper_xaxis))     		  
-    abline(h=seq(0,sp_yaxis,by=0.1),v=c(1:scupper_xaxis)*interval_unit,col="gray")
-  } else if(allele_ratio=="ratio"){
-    plot(chr_dat$POS,chr_dat$RATIO,cex=0.60,xlim=c(0,scupper_xval),ylim=c(0,sp_yaxis),main=paste("Chr",chromind," Variant Discovery Mapping",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Ratio of Variant Reads/Total Reads [ratio]',pch=10, col=plot,xaxt='n')
-    try(lines(loess.smooth(chr_dat$POS,chr_dat$RATIO,span=loess_span),lwd=5,col=loess_color))
-    #plot loess curve for data without subtraction of exclusion variants
-    if(length(exclusion_list)>0){
-      try(lines(loess.smooth(chr_origdat$POS,chr_origdat$adj_AB,span=loess_span),lwd=5,col=excl_loess_color))
-    }
-    axis(1,at=seq(0,scupper_xval,by=interval_unit),labels=c(0:scupper_xaxis))     		  
-    abline(h=seq(0,sp_yaxis,by=0.1),v=c(1:scupper_xaxis)*interval_unit,col="gray")
-  }
-}
-
-######################
-#graph barplots
-location_index<-NULL
-meanSNP_dat<-NULL
-# prepare table of counts and calculations
-for(chromind in interval_frame$CHROM){
-  #for standardized x-axis
-  if(xaxis_standard==TRUE){
-    intervalind<-max(interval_frame$INTERVAL)*1000000/bin_size
-  } else if(xaxis_standard==FALSE){
-    intervalind<-interval_frame$INTERVAL[interval_frame$CHROM==chromind]*1000000/bin_size
-  }
-  #start intervals with **1 and end with **0
-  interval_begin<-c(((0:(intervalind-1))*bin_size)+1)
-  interval_end<-c((1:intervalind)*bin_size)
-  
-  #define x-axis upper limit
-  if(xaxis_standard==TRUE){
-    upper_xaxis<-max(interval_frame$INTERVAL)
-  } else if(xaxis_standard==FALSE){
-    upper_xaxis<-interval_frame$INTERVAL[interval_frame$CHROM==chromind]
-  }
-  #prepare columns
-  snp_counter<-0
-  purealt_counter<-0
-  pureref_counter<-0
-  het_counter<-0
-  chr_mean<-0
-  normed_freq<-0
-  ratio<-0
-  
-  interval_index<-data.frame(chromind,interval_begin,interval_end,snp_counter,purealt_counter,pureref_counter,het_counter,chr_mean,normed_freq)
-  chr_dat<-subset(vcfinfo_dat,CHROM==chromind)
-  #ratio calculation
-  ratio<-chr_dat$AO/(chr_dat$AO+chr_dat$RO)
-  
-  #if counter based on adj_AB or ratio
-  if(allele_ratio=="ratio"){
-    chr_purealtdat<-subset(chr_dat,ratio>=threshold_upper)#;chr_purealtdat
-    chr_purerefdat<-subset(chr_dat,ratio<=threshold_lower)#;chr_purerefdat
-    chr_hetdat<-subset(chr_dat,ratio>threshold_lower & ratio<threshold_upper)#;chr_hetdat
-  } else if(allele_ratio=="AB"){
-    chr_purealtdat<-subset(chr_dat,adj_AB>=threshold_upper)#;chr_purealtdat
-    chr_purerefdat<-subset(chr_dat,adj_AB<=threshold_lower)#;chr_purerefdat
-    chr_hetdat<-subset(chr_dat,adj_AB>threshold_lower & adj_AB<threshold_upper)#;chr_hetdat
-  }
-  #if chromosome with data, count number of snps within each bin (positions rounded up to nearest bin), else skip to next chromosome
-  if(dim(chr_dat)[1]>0){
-    for(i in 1:dim(chr_dat)[1]){
-      chr_datind<-chr_dat[i,]
-      #round up to nearest bin-size interval			
-      chr_datind_upper<-ceiling(chr_datind$POS/bin_size)*bin_size
-      interval_coln<-NULL;interval_rown<-NULL
-      #identify row and and counter column to increment
-      interval_coln<-which(names(interval_index)=="snp_counter")
-      interval_rown<-match(chr_datind_upper,interval_index$interval_end)		
-      interval_index[interval_rown,interval_coln]<-c(interval_index$snp_counter[interval_rown]+1)
-    }
-  }else{
-    next
-  }
-  ##if chromosome with pure AO, count number of snps with each bin (positions rounded up to nearest bin)
-  if(dim(chr_purealtdat)[1]>0){
-    for(i in 1:dim(chr_purealtdat)[1]){
-      chr_purealtind<-chr_purealtdat[i,]
-      chr_purealtind_upper<-ceiling(chr_purealtind$POS/bin_size)*bin_size
-      interval_coln<-NULL;interval_rown<-NULL
-      interval_coln<-which(names(interval_index)=="purealt_counter")
-      interval_rown<-match(chr_purealtind_upper,interval_index$interval_end)		
-      interval_index[interval_rown,interval_coln]<-c(interval_index$purealt_counter[interval_rown]+1)
-    }
-  }
-  #if chromosome with pure RO, count number of snps with each bin (positions rounded up to nearest bin)
-  if(dim(chr_purerefdat)[1]>0){
-    for(i in 1:dim(chr_purerefdat)[1]){
-      chr_purerefind<-chr_purerefdat[i,]
-      chr_purerefind_upper<-ceiling(chr_purerefind$POS/bin_size)*bin_size
-      interval_coln<-NULL;interval_rown<-NULL
-      interval_coln<-which(names(interval_index)=="pureref_counter")
-      interval_rown<-match(chr_purerefind_upper,interval_index$interval_end)			
-      interval_index[interval_rown,interval_coln]<-c(interval_index$pureref_counter[interval_rown]+1)
-    }
-  }
-  #if chromosome with hets, count number of snps with each bin (positions rounded up to nearest bin)
-  if(dim(chr_hetdat)[1]>0){
-    for(i in 1:dim(chr_hetdat)[1]){
-      chr_hetind<-chr_hetdat[i,]
-      chr_hetind_upper<-ceiling(chr_hetind$POS/bin_size)*bin_size
-      interval_coln<-NULL;interval_rown<-NULL
-      interval_coln<-which(names(interval_index)=="het_counter")
-      interval_rown<-match(chr_hetind_upper,interval_index$interval_end)			
-      interval_index[interval_rown,interval_coln]<-c(interval_index$het_counter[interval_rown]+1)
-    }
-  }
-  #irrespective of standardized x-axis, mean should be calculated from actual interval range of chromosome
-  chr_mean<-sum(interval_index$purealt_counter)/(interval_frame$INTERVAL[interval_frame$CHROM==chromind]*1000000/bin_size)
-  interval_index$chr_mean<-chr_mean
-  meanSNP_dat<-rbind(meanSNP_dat,data.frame(chromind,chr_mean))
-  #normalization treatment for if SNPs are AO=0, AO=total SNPs, or AO and RO in bin
-  for(i in 1:dim(interval_index)[1]){
-    chr_intind<-interval_index[i,]
-    #hom definition based on specified upper and lower thresholds
-    if(chr_intind$purealt_counter<=threshold_lower){
-      interval_coln<-NULL
-      interval_coln<-which(names(interval_index)=="normed_freq")
-      interval_index[i,interval_coln]=0
-    } else if (chr_intind$purealt_counter==chr_intind$snp_counter){
-      interval_coln<-NULL
-      interval_coln<-which(names(interval_index)=="normed_freq")
-      interval_index[i,interval_coln]=(chr_intind$purealt_counter)^2/chr_mean
-    } else {
-      interval_coln<-NULL
-      interval_coln<-which(names(interval_index)=="normed_freq")
-      interval_index[i,interval_coln]=chr_mean*(chr_intind$purealt_counter)^2/(chr_intind$snp_counter-chr_intind$purealt_counter)
-    }
-  }
-  location_index<-rbind(location_index,interval_index)
-}
-
-for(chromind in interval_frame$CHROM){
-  interval_index<-location_index[location_index$chromind==chromind,]
-  #assign 0 values to avoid empty datatable error
-  if(dim(interval_index)[1]==0){
-    interval_index[1,]<-rep(0,dim(interval_index)[2])		
-  }
-}
-#set up x_axis 
-if(xaxis_standard==TRUE){
-  #for standardized x-axis (max x-axis chromosome length)
-  bpupper_xaxis<-max(interval_frame$INTERVAL)
-  bpupper_xval<-bpupper_xaxis*interval_unit
-} else if(xaxis_standard==FALSE){
-  bpupper_xaxis<-intervalind
-  bpupper_xval<-intervalind*interval_unit
-}
-#set up y_axis range for barplots
-if(bfreq_norm==TRUE){
-  bp_yaxis<-5*ceiling(max(location_index$normed_freq)/5)
-  #assign non-0 value to yaxis to avoid error
-  if(bp_yaxis==0){
-    bp_yaxis<-10
-  }
-  # }
-  if(xaxis_standard==TRUE){
-    bplot<-barplot(interval_index$normed_freq,space=0,ylim=c(0,bp_yaxis),main=paste("Chr",chromind," Variant Only",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Normalised Frequency')	
-  }else if(xaxis_standard==FALSE){
-    bplot<-barplot(interval_index$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')	
-  }
-}else if(bfreq_norm==FALSE){
-  bp_yaxis<-5*ceiling(max(location_index$purealt_counter)/5)
-  #assign non-0 value to yaxis to avoid error
-  if(bp_yaxis==0){
-    bp_yaxis<-10
-  }
-  # }
-  if(xaxis_standard==TRUE){
-    bplot<-barplot(interval_index$purealt_counter,space=0,ylim=c(0,bp_yaxis),main=paste("Chr",chromind," Variant Only",sep=""),xlab="Position along Chromosome (in Mb)",ylab='Frequency')	
-  }else if(xaxis_standard==FALSE){
-    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')	
-  }
-  
-  bp_xaxis1<-as.numeric(bplot)
-  bp_xaxis2<-c(bp_xaxis1,tail(bp_xaxis1,1)+bp_xaxis1[2]-bp_xaxis1[1])
-  bp_xaxis<-bp_xaxis2-bp_xaxis1[1]
-  
-  axis(1,at=bp_xaxis,labels=seq(0,bpupper_xaxis,by=c(bin_size/1000000)))	
-}
-dev.off()
-
-