# HG changeset patch # User pmac # Date 1464767572 14400 # Node ID 087ab474dfa477451b47781bf54f89c78923ef22 Uploaded diff -r 000000000000 -r 087ab474dfa4 normalizeMedianNegCntrl.pl --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/normalizeMedianNegCntrl.pl Wed Jun 01 03:52:52 2016 -0400 @@ -0,0 +1,87 @@ +############################################################################### +# In summary this script normalizes each plate of input data to each of its negative +# controls. +# +# Args: +# input.data.frame contains one or more columns of plate data. The first column +# must contain the well annotation ie A01, A02 etc. Each subsequent column represents +# a plate of data where the column name is the plate name. +# +# plate.conf.data.frame is a file of the following format: +# well, type (syntax must be either "poscontr" or "negcontr" or "empty"), name (name of control) +# +# Returns: +# plate values normalized to each negative control type per plate. +# +# Author: kate gould +############################################################################### + +use strict; +use warnings; +use IO::Handle; +use File::Temp qw/ tempfile tempdir /; +my $tdir = tempdir( CLEANUP => 0 ); + +# check to make sure having correct input and output files +my $usage = "usage: normalizeMedianNegCntrl.pl [TABULAR.in] [TABULAR.in] [TABULAR.out] \n"; +die $usage unless @ARGV == 3; + +#get the input arguments +my $plateData = $ARGV[0]; +my $plateConfig = $ARGV[1]; +my $normMedNeg = $ARGV[2]; + +#open the input files +open (INPUT1, "<", $plateData) || die("Could not open file $plateData \n"); +open (INPUT2, "<", $plateConfig) || die("Could not open file $plateConfig \n"); +open (OUTPUT1, ">", $normMedNeg) || die("Could not open file $normMedNeg \n"); + +#variable to store the name of the R script file +my $r_script; + +# R script to implement the calcualtion of q-values based on multiple simultaneous tests p-values +# construct an R script file and save it in a temp directory +chdir $tdir; +$r_script = "normalizeMedianNegCntrl.r"; + +open(Rcmd,">", $r_script) or die "Cannot open $r_script \n\n"; +print Rcmd " + #options(show.error.messages = FALSE); + header <- read.table(\"$plateData\", head=F, sep=\"\\t\", comment=\"\", nrows=1); + input.data.frame <- read.table(\"$plateData\", head=F, sep=\"\\t\", skip=1); + plate.conf.data.frame <- read.table(\"$plateConfig\", head=T, sep=\"\\t\", comment=\"\"); + + normalise <- function(x, y){if (!is.na(x)) return (x/y) else (return (NA))} + + # assumed second column is type ie negcontr and third column is name of control + negative.controls.list <- unique(plate.conf.data.frame[which (plate.conf.data.frame[,2] == \"negcontr\"),][,3]); + + normalized.data.frame <- data.frame(well=input.data.frame[,1], stringsAsFactors=FALSE); + column.name.list <- c(); + for (negative.control in negative.controls.list){ + negative.control.wells <- plate.conf.data.frame[which (plate.conf.data.frame[3] == negative.control),][,1]; + + for (i in 2:length(colnames(input.data.frame))){ + column.name <- paste(colnames(input.data.frame)[i], \".normalised.to.\", negative.control, sep=\"\"); + column.name.list <- append(column.name.list, column.name); + + negative.control.values <- input.data.frame[((input.data.frame[,1] %in% negative.control.wells)&(!(is.na(input.data.frame[,i])))),][,i]; + median.negative.value <- median(negative.control.values); + + normalised.to.median.negative.control.value <- round(sapply(input.data.frame[,i], FUN=normalise, median.negative.value), 2); + normalized.data.frame <- cbind(normalized.data.frame, normalised.to.median.negative.control.value); + } + } + header <- c(header[1], column.name.list) + write.table(header, file=\"$normMedNeg\", quote=F, sep=\"\\t\", row.names=F, col.names=F); + write.table(normalized.data.frame, file=\"$normMedNeg\", quote=F, sep=\"\\t\", row.names=F, col.names=F, append=T); + #eof\n"; + +close Rcmd; + +system("R --no-restore --no-save --no-readline < $r_script > $r_script.out"); + +#close the input and output files +close(OUTPUT1); +close(INPUT1); +close(INPUT2); diff -r 000000000000 -r 087ab474dfa4 normalizeMedianNegCntrl.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/normalizeMedianNegCntrl.xml Wed Jun 01 03:52:52 2016 -0400 @@ -0,0 +1,135 @@ + + + + normalizeMedianNegCntrl.pl $inputFile1 $inputFile2 $outputFile1 + + + + + + + + + + + + + + + + + + + + + +.. class:: infomark + +**What it does** + +This program takes a linear plate table and a plate config file as input and then normalizes each plate in the table to the negative controls in the config. +The plate config file **must** contain the columns Well, Type and Name in that order. Further the data in column Type must be **exclude**, **negcontr** or **poscontr**. + +**Example** + +If the plates table consisted of:: + + #Well Table_1 Table_2 A01 1654 6 A02 1719 8 A03 1624 19 A04 1518 6 A05 1587 12 A06 1517 8 A07 1638 9 A08 1733 7 A09 1617 6 A10 1722 8 ... + P14 1630 6 P15 1576 6 P16 1740 7 P17 1578 6 P18 1544 6 P19 1692 6 P20 1560 6 P21 1552 6 P22 1790 7 P23 1537 6 P24 NA NA + +and the plates config was:: + + #Well Type Name A02 negcontr NT2 + B02 negcontr NT2 + C02 poscontr ASCIZ + D02 poscontr PARP + E02 negcontr NT2 + F02 poscontr ASCIZ + G02 poscontr PARP + H02 negcontr NT2 + I02 poscontr ASCIZ + J02 poscontr PARP + K02 negcontr NT2 + L02 poscontr ASCIZ + M02 poscontr PARP + N02 negcontr NT2 + O02 poscontr ASCIZ + P02 negcontr NT2 + A23 negcontr NT2 + B23 negcontr NT2 + C23 poscontr ASCIZ + D23 poscontr PARP + E23 negcontr NT2 + F23 poscontr ASCIZ + G23 poscontr PARP + H23 negcontr NT2 + I23 poscontr ASCIZ + J23 poscontr PARP + K23 negcontr NT2 + L23 poscontr ASCIZ + M23 poscontr PARP + N23 negcontr NT2 + O23 poscontr ASCIZ + P23 negcontr NT2 + A01 exclude exclude + A24 exclude exclude + B01 exclude exclude + B24 exclude exclude + C01 exclude exclude + C24 exclude exclude + D01 exclude exclude + D24 exclude exclude + E01 exclude exclude + E24 exclude exclude + F01 exclude exclude + F24 exclude exclude + G01 exclude exclude + G24 exclude exclude + H01 exclude exclude + H24 exclude exclude + I01 exclude exclude + I24 exclude exclude + J01 exclude exclude + J24 exclude exclude + K01 exclude exclude + K24 exclude exclude + L01 exclude exclude + L24 exclude exclude + M01 exclude exclude + M24 exclude exclude + N01 exclude exclude + N24 exclude exclude + O01 exclude exclude + O24 exclude exclude + P01 exclude exclude + P24 exclude exclude + +Running the program will give the following output:: + + #Well Table_1 Table_2 + A01 1.03 0.75 + A02 1.07 1 + A03 1.02 2.38 + A04 0.95 0.75 + A05 0.99 1.5 + A06 0.95 1 + A07 1.02 1.12 + A08 1.08 0.88 + A09 1.01 0.75 + A10 1.08 1 + ... + P14 1.02 0.75 + P15 0.99 0.75 + P16 1.09 0.88 + P17 0.99 0.75 + P18 0.97 0.75 + P19 1.06 0.75 + P20 0.98 0.75 + P21 0.97 0.75 + P22 1.12 0.88 + P23 0.96 0.75 + P24 NA NA + + + + diff -r 000000000000 -r 087ab474dfa4 test-data/150615-HR-12015-01A.linear --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/test-data/150615-HR-12015-01A.linear Wed Jun 01 03:52:52 2016 -0400 @@ -0,0 +1,385 @@ +#Well Table_1 Table_2 Table_3 Table_4 Table_5 Table_6 +A01 2215 8.88 55.62 2.57 2.91 78.76 +A02 2811 8.61 57.35 1.07 2.03 63.32 +A03 2226 10.96 70.4 0.54 2.41 79.41 +A04 2528 9.35 61.59 1.66 2.14 74.9 +A05 1827 11.87 73.78 0.66 2.27 79.31 +A06 2337 8.69 58.49 1.07 1.87 62.34 +A07 2233 9.7 63.41 1.07 2.32 77.48 +A08 2462 8.48 57.39 1.06 2.07 67.32 +A09 2790 9.5 62.83 1.65 2.52 80.57 +A10 2792 9.19 60.67 1.9 2.38 73.6 +A11 2547 9.51 61.76 1.33 2.8 84.6 +A12 2629 9.47 60.94 2.09 2.42 72.34 +A13 1655 11.8 71.24 0.85 2.3 71.88 +A14 2647 8.27 52.13 2.15 2.37 72.76 +A15 2300 11.5 72.17 0.57 2.68 86.26 +A16 2030 11.71 71.63 0.74 2.66 89.03 +A17 2124 9.78 63.89 1.13 2.45 81.13 +A18 2461 9.14 59.65 1.83 2.3 77.14 +A19 2273 8.68 57.85 0.97 2.32 69.57 +A20 2281 10.62 70.58 0.7 1.98 65.3 +A21 2412 10.49 69.86 0.46 2.07 70.27 +A22 1331 10.72 69.65 0.68 1.97 61.54 +A23 2245 9.88 66.24 0.4 2.17 66.33 +A24 2037 8.13 53.26 1.42 2.06 70.05 +B01 2184 10.31 65.29 0.87 2.33 72.4 +B02 2104 9.33 57.94 3.09 2.44 79.26 +B03 1639 11.13 69.68 0.55 2.03 67.42 +B04 2467 9.84 64.33 0.77 2.03 67.95 +B05 1818 11.75 75.25 0.66 1.97 68.96 +B06 1769 9.08 57.32 2.43 2.29 69.18 +B07 1654 12.22 75.7 0.6 2.14 75.29 +B08 2199 10.04 68.58 0.73 2.43 73.55 +B09 2043 8.74 59.03 1.22 1.96 57.65 +B10 1329 11.6 73.06 0.68 2.04 58.75 +B11 2533 7.31 47.06 3.83 3.01 95.3 +B12 2712 9.31 61.98 2.06 2.49 75.2 +B13 1824 10.83 68.42 1.54 2.45 80.09 +B14 2718 7.68 49.74 4.01 2.54 81.37 +B15 2443 10.4 68.81 1.02 2.64 91.33 +B16 2061 11.82 77.1 0.39 2.53 87.74 +B17 2088 10.83 70.69 0.62 2.22 74.12 +B18 2730 10.36 70.4 0.81 2.39 78.69 +B19 2279 8.09 54.28 1.14 1.98 61.28 +B20 2371 12.37 78.28 0.25 2.23 78.36 +B21 2114 10.49 71.85 0.85 2.32 73.66 +B22 2355 9.84 66.71 0.72 2.04 64.63 +B23 1154 11.75 72.1 0.35 2.35 65.16 +B24 1646 7.49 50.12 1.82 2.11 64.74 +C01 2356 9.67 64.43 1.02 2.07 61.55 +C02 2680 10.27 67.57 0.82 2.03 60.64 +C03 2305 8.51 55.88 1.56 1.87 56.38 +C04 2510 9.55 63.55 1 2.02 58.22 +C05 2650 10.23 67.96 0.3 1.95 59.48 +C06 2918 9.76 64.98 0.69 2.21 65.35 +C07 2618 8.28 55.39 1.26 2.42 68.98 +C08 2654 9.56 65.3 1.09 2.52 75.17 +C09 2828 8.44 57.39 1.34 2.21 63.59 +C10 2877 9.48 64.75 1.32 2.59 77.38 +C11 2441 9.78 65.38 1.47 2.38 73.63 +C12 2870 7.1 47.04 3.69 2.87 88.54 +C13 3202 9.02 59.93 1.72 2.43 78.2 +C14 2619 8.63 57.43 2.1 2.93 95.11 +C15 2716 8.82 58.73 1.18 2.46 78.45 +C16 2413 10.72 71.07 0.33 2.62 84.52 +C17 2893 8.18 56.14 1.52 2.8 83.32 +C18 2669 9.82 66.62 0.67 2.26 70.88 +C19 2272 10.81 71.79 0.79 2.1 68.8 +C20 2161 9.42 63.4 1.2 2.79 79.7 +C21 2677 8.75 59.62 0.78 2.04 62.52 +C22 2406 9.91 65 0.71 2.08 68.61 +C23 2494 7.54 49.44 2.33 2.25 63.14 +C24 1760 7.85 51.99 1.82 2.15 66.92 +D01 2206 7.2 45.01 2.77 1.41 36.44 +D02 1033 10.08 62.25 1.84 2.61 77.42 +D03 1744 7.58 49.14 3.04 2.4 70.59 +D04 2579 8.78 59.83 1.2 2.12 62.68 +D05 1921 9.41 64.08 0.94 2.29 70.86 +D06 2352 9.68 63.95 2.3 2.6 79.33 +D07 1844 8.86 59.06 2.11 2.76 85.21 +D08 2831 8.45 56.55 1.77 2.62 78 +D09 1803 9.59 63.67 1.22 2.81 84.73 +D10 2531 9.03 59.9 1.78 2.64 75.75 +D11 1942 7.37 48.51 3.24 3.1 95.33 +D12 2653 9.72 66.34 1.43 2.77 84.49 +D13 2624 6.77 43.52 4.54 3.06 101.35 +D14 2777 6.68 43.03 5.47 3.36 106.95 +D15 2523 8.53 58.7 1.43 3.13 101.22 +D16 2532 9.9 66.11 0.87 2.96 89.01 +D17 2243 9.92 66.79 0.76 3.15 105.74 +D18 2653 11.31 76.1 0.45 2.67 85.7 +D19 1806 11.24 73.26 0.44 2.77 90.06 +D20 2502 11.17 74.5 0.48 2.58 76.4 +D21 2355 9.81 67.22 0.85 2.69 88.68 +D22 2036 11.95 77.65 0.29 2.54 82.78 +D23 4 19.75 100 0 1.5 74.25 +D24 2056 7.06 45.82 1.9 2.27 76.56 +E01 1953 7.89 52.02 2.36 2.09 64.14 +E02 58 10.38 67.24 6.9 2.34 81.33 +E03 1429 11 70.89 0.91 2.09 65.5 +E04 2419 9.41 61.06 1.2 2.4 69.87 +E05 2326 9.41 62.64 1.25 2.36 69.72 +E06 2281 9.32 61.03 2.59 2.88 89.16 +E07 2524 7.92 52.61 2.73 2.63 78 +E08 2444 9.17 62.19 1.51 2.8 84.21 +E09 2402 9.13 60.95 1.5 2.6 78.8 +E10 2470 9.23 60.89 1.94 2.64 78.64 +E11 2387 9.57 63.3 1.59 2.58 80.7 +E12 2812 9.54 64.76 1.21 2.61 83.54 +E13 2121 9.11 60.68 2.22 2.37 70.04 +E14 2467 8.67 58.25 1.34 2.65 87.2 +E15 1932 6.3 39.23 5.64 2.89 91.38 +E16 2207 9.54 63.57 1.54 2.63 86.13 +E17 866 11.03 69.98 0.81 2.23 71.41 +E18 1665 8.31 55.56 2.34 3.14 93.34 +E19 1668 11.19 72.96 0.66 2.3 73.16 +E20 1942 10.59 70.19 0.46 2.46 76.69 +E21 2717 10.46 69.82 0.44 2.22 72.24 +E22 1995 9.92 67.27 1.45 2.73 86.76 +E23 2686 8.98 60.65 1.19 2.33 71.8 +E24 1711 8.25 53.71 1.81 2.43 78.22 +F01 2647 8.61 56.89 1.44 2.05 56.86 +F02 1866 10.14 66.4 0.91 1.86 55.02 +F03 2327 7.3 46.41 4.08 2.04 60.29 +F04 1831 6.3 39.16 6.66 2.25 67.66 +F05 2081 8.89 57.33 2.4 2.34 70.01 +F06 2360 9.29 62.12 2.54 2.41 77.11 +F07 2465 8.21 54.36 2.92 2.59 77.73 +F08 2561 7.36 49 2.89 2.31 66.52 +F09 2242 9.81 66.77 1.43 2.47 72.7 +F10 2595 6.08 37.5 6.2 2.39 68.73 +F11 2601 8.19 54.48 3.65 2.77 96.56 +F12 2075 9.72 63.04 1.59 2.47 72.8 +F13 2452 9.25 62.11 1.51 2.66 82.33 +F14 2506 8.22 53.79 2.04 2.54 77.56 +F15 2228 8.19 53.82 2.65 2.91 103.54 +F16 2869 7.83 52.49 1.92 2.58 76.14 +F17 1322 8.78 60.14 1.97 2.52 72.6 +F18 2712 7.56 50 3.39 2.68 81.09 +F19 2623 9 60.62 1.03 2.41 73.71 +F20 2320 9.37 62.5 1.51 2.43 70.9 +F21 2528 9.04 59.53 1.15 2.2 65.33 +F22 1797 6.44 39.96 4.67 2.38 70.58 +F23 1933 10.26 67.36 0.93 2.5 71.62 +F24 1759 8.4 56.17 1.99 1.92 67.93 +G01 2715 10.83 72.08 0.59 1.98 58.65 +G02 2567 11.22 71.76 0.51 1.95 59.34 +G03 2346 8.73 57.37 1.49 2.13 61.93 +G04 2536 7.29 46.77 4.89 2.24 65.72 +G05 2157 8.33 54.38 2.23 2.35 65.99 +G06 2549 6.64 41.66 5.1 2.59 78.36 +G07 2368 9.22 61.15 1.35 2.41 73.63 +G08 2798 9.61 63.44 1.18 2.46 72.8 +G09 2595 8.97 59.73 1.93 2.4 68.47 +G10 3309 6.57 42.16 3.84 2.34 62.92 +G11 2701 7.84 53.05 2.3 2.46 68.96 +G12 2916 7.37 49.04 3.19 2.61 72.96 +G13 2664 9.26 63.48 1.73 2.88 88.03 +G14 2998 7.58 51.17 2.94 2.6 75.25 +G15 2257 9.19 63.14 1.2 2.49 71.58 +G16 2850 10.29 70.53 1.09 2.33 71.99 +G17 2559 7.88 52.25 2.34 2.67 83.35 +G18 2684 8.49 55.48 2.72 2.69 83.61 +G19 2852 7.68 50.25 3.54 2.78 87.58 +G20 2779 7.11 45.88 3.17 2.58 73.2 +G21 2448 9.83 67.28 0.69 2.52 77.41 +G22 2741 7.4 48.23 3.1 2.42 68.04 +G23 2945 10.08 70.12 0.75 2.52 78.27 +G24 1740 6.25 38.91 3.33 2.19 65.21 +H01 2214 12.71 78.73 0.36 2.1 68.06 +H02 11 19.73 90.91 0 2.18 67.36 +H03 1344 10.4 65.77 1.86 2.36 69.92 +H04 1937 7.72 49.3 3.1 2.42 70.48 +H05 2130 9.08 60.19 1.41 2.39 67.18 +H06 2637 8.69 56.16 1.71 2.34 68.97 +H07 2278 10.21 67.08 1.14 2.85 85.95 +H08 2020 10.78 72.62 1.09 2.56 74.15 +H09 1780 10.32 67.25 1.91 3.05 82.88 +H10 2262 7.84 52.43 2.25 3.01 78.92 +H11 2130 12.63 80.47 0.47 3.17 96.52 +H12 2404 9.57 63.89 1.87 3.07 87.48 +H13 1872 10.9 70.78 1.23 3.15 91.34 +H14 2598 9.23 63.01 1.31 2.65 76.86 +H15 2164 10.57 72.87 1.06 2.74 78.64 +H16 2475 9.77 66.87 0.73 2.86 90.95 +H17 1783 11.15 72.74 0.67 2.97 88.69 +H18 2114 9.08 60.83 1.32 2.87 84.31 +H19 2621 7.59 49.83 3.13 2.69 81.29 +H20 2623 7.95 53.49 2.06 2.34 68.36 +H21 1954 10.2 66.27 1.64 2.5 78.56 +H22 2356 9.53 62.82 1.57 3.31 99.15 +H23 8 26 100 0 3.25 139 +H24 1421 7.88 47.64 4.93 1.98 78.52 +I01 2071 12.39 78.17 0.43 2.11 61.97 +I02 1366 12.04 75.62 0.88 2.36 65.33 +I03 1769 9.91 63.2 1.19 2.13 56.36 +I04 2389 10.75 70.24 0.63 2.28 65.29 +I05 1983 8 51.49 2.57 2.69 74.38 +I06 2209 9.97 64.46 1.45 2.9 80.24 +I07 2524 9.56 62.08 1.78 3.01 88.14 +I08 1961 11.84 75.68 0.66 3.23 88.78 +I09 1925 11.12 71.27 0.88 2.82 78.49 +I10 2364 10.85 69.42 0.89 3.17 89.68 +I11 1809 12.46 77.34 0.44 2.66 76.47 +I12 2387 10.95 70.38 0.88 3.15 87.18 +I13 2530 12.5 78.42 0.87 3.04 94.74 +I14 2086 12.23 75.6 0.91 3.47 101.93 +I15 2587 10.9 72.59 0.7 2.84 85.07 +I16 2480 12.24 78.15 0.44 3.45 108.16 +I17 1770 12.3 78.25 0.73 3.12 95.09 +I18 1964 8.3 55.09 2.9 3.32 96.99 +I19 1969 10.6 69.78 1.12 3.11 95.42 +I20 2330 8.19 53.22 2.1 3.43 100.06 +I21 1922 8.76 58.27 1.82 3.08 90.8 +I22 2062 11.61 75.12 0.58 3.06 95.76 +I23 2446 9.95 66.39 1.19 2.99 83.27 +I24 1457 9.09 57.79 2.2 3.49 102.71 +J01 1967 13.45 80.22 0.31 2.06 62 +J02 1954 11.02 71.65 1.23 2.08 57.26 +J03 1402 10.14 67.05 0.86 1.75 48.25 +J04 1316 10.69 70.82 0.68 2.08 59.11 +J05 1784 7 45.8 4.37 2.09 58.3 +J06 2557 7.78 52.33 2.66 2.73 77.66 +J07 1753 10.05 66.29 1.43 2.44 71.51 +J08 1844 11.57 75.43 0.98 2.59 73.92 +J09 1670 9.2 61.44 0.78 2.5 65.14 +J10 2427 9.55 64.19 1.03 2.68 75.03 +J11 2011 11.93 77.03 0.2 2.55 73.74 +J12 1973 11.51 74.71 0.91 2.92 82.11 +J13 2276 11.43 74.74 0.66 3 89.39 +J14 2088 11.24 74.47 0.67 2.94 87.12 +J15 2476 10.48 69.35 0.65 2.77 79.58 +J16 2395 10.95 71.9 0.92 3.22 94.31 +J17 2127 11.9 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PLK 5347 K23 poscontr Hamp 57817 L23 negcontr OTP -1 M23 poscontr MPEG1 17476 N23 negcontr OTP -1 O23 negcontr mock -1 \ No newline at end of file