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     1 ###############################################################################
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     2 # In summary this script calculates the z prime factor for one or more plates
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     3 # of data.
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     4 #
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     5 # Args:
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     6 #   input.data.frame contains one or more columns of plate data. The first column 
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     7 #   must contain the well annotation ie A01, A02 etc. Each subsequent column represents
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     8 #   a plate of data where the column name is the plate name.
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     9 #
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    10 #   plate.conf.data.frame is a file of the following format:
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    11 #	  well, type(syntax must be either "poscontr" or "negcontr"), name (name of control)
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    12 #
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    13 # Returns:
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    14 #    z'factor for each plate for each combination of each type of positive vs negative control.
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    15 #
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    16 # Author: gouldkate
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    17 ###############################################################################
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    18 
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    19 use strict;
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    20 use warnings;
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    21 use IO::Handle;
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    22 use File::Temp qw/ tempfile tempdir /;
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    23 my $tdir = tempdir( CLEANUP => 0 );
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    24 
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    25 # check to make sure having correct input and output files
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    26 my $usage = "usage: calculateZPrimeFactor.pl [TABULAR.in] [TABULAR.in] [TABULAR.out] \n";
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    27 die $usage unless @ARGV == 3;
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    28 
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    29 #get the input arguments
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    30 my $plateData = $ARGV[0];
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    31 my $plateConfig =  $ARGV[1];
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    32 my $zPrimeFactor =  $ARGV[2];
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    33 
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    34 #open the input files
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    35 open (INPUT1, "<", $plateData) || die("Could not open file $plateData \n");
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    36 open (INPUT2, "<", $plateConfig) || die("Could not open file $plateConfig \n");
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    37 open (OUTPUT1, ">", $zPrimeFactor) || die("Could not open file $zPrimeFactor \n");
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    38 
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    39 #variable to store the name of the R script file
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    40 my $r_script;
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    41 
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    42 # R script to implement the calcualtion of q-values based on multiple simultaneous tests p-values 	
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    43 # construct an R script file and save it in a temp directory
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    44 chdir $tdir;
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    45 $r_script = "calculateZPrimeFactor.r";
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    46 
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    47 open(Rcmd,">", $r_script) or die "Cannot open $r_script \n\n"; 
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    48 print Rcmd "
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    49 	#options(show.error.messages = FALSE);
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    50 	input.data.frame <- read.table(\"$plateData\", head=T, sep=\"\\t\", comment=\"\");
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    51 	plate.conf.data.frame <- read.table(\"$plateConfig\", head=T, sep=\"\\t\", comment=\"\");
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    52 	
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    53 	# assumed second column is type ie negcontr or poscontr and third column is name of control
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    54 	negative.controls.list <- unique(plate.conf.data.frame[which (plate.conf.data.frame[,2] == \"negcontr\"),][,3])
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    55 	positive.controls.list <- unique(plate.conf.data.frame[which (plate.conf.data.frame[,2] == \"poscontr\"),][,3])
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    56   
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    57 	z.prime.factor.report.data.frame <- data.frame()
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    58   
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    59   
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    60 	for (negative.control in negative.controls.list){
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    61 		for (positive.control in positive.controls.list){
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    62 			for (i in 2:length(colnames(input.data.frame))){
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    63         
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    64 				negative.control.wells <- plate.conf.data.frame[which (plate.conf.data.frame[3] == negative.control),][,1]
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    65 				positive.control.wells <- plate.conf.data.frame[which (plate.conf.data.frame[3] == positive.control),][,1]
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    66 
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    67 				control.duo <- paste(negative.control, positive.control, sep=\"/\")
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    68 				
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    69 				negative.control.values <- input.data.frame[((input.data.frame[,1] %in% negative.control.wells)&(!(is.na(input.data.frame[,i])))),][,i]
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    70 				positive.control.values <- input.data.frame[((input.data.frame[,1] %in% positive.control.wells)&(!(is.na(input.data.frame[,i])))),][,i]
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    71 
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    72 				if ((length(negative.control.values)==0)|(length(positive.control.values)==0)){
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    73 					z.prime.factor <- NA
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    74 				} else {
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    75 					if ((sum(negative.control.values)) < (sum(positive.control.values))){
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    76 						low.value.controls <- negative.control.values
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    77 						high.value.controls <- positive.control.values
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    78 					} else {
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    79 						low.value.controls <- positive.control.values
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    80 						high.value.controls <- negative.control.values
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    81 					}
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    82 					z.prime.factor <- round(1 - ((3 * (sd(high.value.controls, na.rm = TRUE) + sd(low.value.controls, na.rm = TRUE))/(abs(mean(high.value.controls, na.rm = TRUE) - mean(low.value.controls, na.rm = TRUE))))), 2)
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    83 				}
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    84 				
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    85 				plate.name <- colnames(input.data.frame)[i]
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    86 				z.prime.factor.current.row.data.frame <- data.frame(plate=plate.name, control.duo=control.duo, z.prime.factor=z.prime.factor, stringsAsFactors=FALSE)
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    87 				z.prime.factor.report.data.frame <-rbind(z.prime.factor.report.data.frame, z.prime.factor.current.row.data.frame)
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    88 			}
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    89 		}
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    90 	}
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    91 	z.prime.factor.report.data.frame <- z.prime.factor.report.data.frame[order(z.prime.factor.report.data.frame\$control.duo),]
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    92 	write.table(z.prime.factor.report.data.frame, file=\"$zPrimeFactor\", quote=F, sep=\"\\t\", row.names=F);
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    93 	#eof\n";
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    94 
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    95 close Rcmd;	
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    96 
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    97 system("R --no-restore --no-save --no-readline < $r_script > $r_script.out");
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    98 
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    99 #close the input and output files
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   100 close(OUTPUT1);
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   101 close(INPUT1);
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   102 close(INPUT2);
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