Limma read GenePix version 6 file
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Ye, Bin ▴ 150
@ye-bin-1280
Last seen 9.7 years ago
Hi, When I try to read the GenePix output file from GenePix 6, I got an error as below: Error in "[.data.frame"(obj, , columns[[a]]) : undefined columns selected I suspect GenePix 6 have some new columns different than previous version: Block Column Row Name ID X Y Dia. F635 Median F635 Mean F635 SD F635 CV B635 B635 Median B635 Mean B635 SD B635 CV % > B635+1SD % > B635+2SD F635 % Sat. F532 Median F532 Mean F532 SD F532 CV B532 B532 Median B532 Mean B532 SD B532 CV % > B532+1SD % > B532+2SD F532 % Sat. Ratio of Medians (635/532) Ratio of Means (635/532) Median of Ratios (635/532) Mean of Ratios (635/532) Ratios SD (635/532) Rgn Ratio (635/532) Rgn R2 (635/532) F Pixels B Pixels Circularity Sum of Medians (635/532) Sum of Means (635/532) Log Ratio (635/532) F635 Median - B635 F532 Median - B532 F635 Mean - B635 F532 Mean - B532 F635 Total Intensity F532 Total Intensity SNR 635 SNR 532 Flags Normalize Autoflag Can anybody help me out? We need to deal with GenePix data for our experiment. Thanks in advance! Bin
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@saroj-mohapatra-1446
Last seen 9.7 years ago
Bin: Which function are you using for reading the data? If you are using the limma function read.maimages, then the function provides some extra arguments that can force it to read specific columns. The help file says: ---- > ?read.maimages() columns: list with fields 'Rf', 'Gf', 'Rb' and 'Gb' giving the column names to be used for red and green foreground and background or, in the case of Imagene data, a list with fields 'f' and 'b'. This argument is not usually specified by the user but, if it is, it over-rides 'source'. ----- Something like this might work: myRG = read.maimages(myGenepix6Files, columns=list(Rf='F635 Median', Gf='F532 Median', Rb = 'B635 Median', Gb='B532 Median'), ...) Also, there are other arguments (other.columns) in the function for reading more columns from the file. Hope it helps. Best, Saroj Ye, Bin wrote: >Hi, > >When I try to read the GenePix output file from GenePix 6, I got an error as below: > >Error in "[.data.frame"(obj, , columns[[a]]) : > undefined columns selected > >I suspect GenePix 6 have some new columns different than previous version: > >Block Column Row Name ID X Y Dia. F635 Median F635 Mean F635 SD F635 CV B635 B635 Median B635 Mean B635 SD B635 CV % > B635+1SD % > B635+2SD F635 % Sat. F532 Median F532 Mean F532 SD F532 CV B532 B532 Median B532 Mean B532 SD B532 CV % > B532+1SD % > B532+2SD F532 % Sat. Ratio of Medians (635/532) Ratio of Means (635/532) Median of Ratios (635/532) Mean of Ratios (635/532) Ratios SD (635/532) Rgn Ratio (635/532) Rgn R2 (635/532) F Pixels B Pixels Circularity Sum of Medians (635/532) Sum of Means (635/532) Log Ratio (635/532) F635 Median - B635 F532 Median - B532 F635 Mean - B635 F532 Mean - B532 F635 Total Intensity F532 Total Intensity SNR 635 SNR 532 Flags Normalize Autoflag > >Can anybody help me out? We need to deal with GenePix data for our experiment. > >Thanks in advance! > > >Bin > >_______________________________________________ >Bioconductor mailing list >Bioconductor at stat.math.ethz.ch >https://stat.ethz.ch/mailman/listinfo/bioconductor > >
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Ye, Bin ▴ 150
@ye-bin-1280
Last seen 9.7 years ago
It works! Thank you, Saroj! Have a good weekend! Bin -----Original Message----- From: Saroj Mohapatra [mailto:smohapat@vbi.vt.edu] Sent: Fri 10/14/2005 3:19 PM To: Ye, Bin Cc: bioconductor at stat.math.ethz.ch Subject: Re: [BioC] Limma read GenePix version 6 file Bin: Which function are you using for reading the data? If you are using the limma function read.maimages, then the function provides some extra arguments that can force it to read specific columns. The help file says: ---- > ?read.maimages() columns: list with fields 'Rf', 'Gf', 'Rb' and 'Gb' giving the column names to be used for red and green foreground and background or, in the case of Imagene data, a list with fields 'f' and 'b'. This argument is not usually specified by the user but, if it is, it over-rides 'source'. ----- Something like this might work: myRG = read.maimages(myGenepix6Files, columns=list(Rf='F635 Median', Gf='F532 Median', Rb = 'B635 Median', Gb='B532 Median'), ...) Also, there are other arguments (other.columns) in the function for reading more columns from the file. Hope it helps. Best, Saroj Ye, Bin wrote: >Hi, > >When I try to read the GenePix output file from GenePix 6, I got an error as below: > >Error in "[.data.frame"(obj, , columns[[a]]) : > undefined columns selected > >I suspect GenePix 6 have some new columns different than previous version: > >Block Column Row Name ID X Y Dia. F635 Median F635 Mean F635 SD F635 CV B635 B635 Median B635 Mean B635 SD B635 CV % > B635+1SD % > B635+2SD F635 % Sat. F532 Median F532 Mean F532 SD F532 CV B532 B532 Median B532 Mean B532 SD B532 CV % > B532+1SD % > B532+2SD F532 % Sat. Ratio of Medians (635/532) Ratio of Means (635/532) Median of Ratios (635/532) Mean of Ratios (635/532) Ratios SD (635/532) Rgn Ratio (635/532) Rgn R2 (635/532) F Pixels B Pixels Circularity Sum of Medians (635/532) Sum of Means (635/532) Log Ratio (635/532) F635 Median - B635 F532 Median - B532 F635 Mean - B635 F532 Mean - B532 F635 Total Intensity F532 Total Intensity SNR 635 SNR 532 Flags Normalize Autoflag > >Can anybody help me out? We need to deal with GenePix data for our experiment. > >Thanks in advance! > > >Bin > >_______________________________________________ >Bioconductor mailing list >Bioconductor at stat.math.ethz.ch >https://stat.ethz.ch/mailman/listinfo/bioconductor > >
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@gordon-smyth
Last seen 1 hour ago
WEHI, Melbourne, Australia
limma has been well tested with GenePix 6 and there should be no problem. Specifying columns explicitly should not be required. If you believe there's a limma error, please give the code you used so that we can debug. BTW, the columns that Saroj specified (all medians) can now be given as source="genepix.median" Gordon >Date: Fri, 14 Oct 2005 16:11:25 -0400 >From: "Ye, Bin" <binye at="" med.wayne.edu=""> >Subject: Re: [BioC] Limma read GenePix version 6 file >To: "Saroj Mohapatra" <smohapat at="" vbi.vt.edu=""> >Cc: bioconductor at stat.math.ethz.ch > >It works! Thank you, Saroj! > >Have a good weekend! > > > >Bin > > >-----Original Message----- >From: Saroj Mohapatra [mailto:smohapat at vbi.vt.edu] >Sent: Fri 10/14/2005 3:19 PM >To: Ye, Bin >Cc: bioconductor at stat.math.ethz.ch >Subject: Re: [BioC] Limma read GenePix version 6 file > >Bin: > >Which function are you using for reading the data? > >If you are using the limma function read.maimages, then the function >provides some extra arguments that can force it to read specific >columns. The help file says: > >---- > > ?read.maimages() > >columns: list with fields 'Rf', 'Gf', 'Rb' and 'Gb' giving the column > names to be used for red and green foreground and background > or, in the case of Imagene data, a list with fields 'f' and > 'b'. This argument is not usually specified by the user but, > if it is, it over-rides 'source'. >----- > >Something like this might work: > >myRG = read.maimages(myGenepix6Files, columns=list(Rf='F635 Median', >Gf='F532 Median', Rb = 'B635 Median', Gb='B532 Median'), ...) > >Also, there are other arguments (other.columns) in the function for >reading more columns from the file. > >Hope it helps. Best, > >Saroj > >Ye, Bin wrote: > > >Hi, > > > >When I try to read the GenePix output file from GenePix 6, I got an > error as below: > > > >Error in "[.data.frame"(obj, , columns[[a]]) : > > undefined columns selected > > > >I suspect GenePix 6 have some new columns different than previous version: > > > >Block Column Row Name ID X Y Dia. F635 > Median F635 Mean F635 SD F635 CV B635 B635 Median B635 > Mean B635 SD B635 CV % > B635+1SD % > B635+2SD F635 % > Sat. F532 Median F532 Mean F532 SD F532 CV B532 B532 > Median B532 Mean B532 SD B532 CV % > B532+1SD % > > B532+2SD F532 % Sat. Ratio of Medians (635/532) Ratio of > Means (635/532) Median of Ratios (635/532) Mean of Ratios > (635/532) Ratios SD (635/532) Rgn Ratio (635/532) Rgn R2 > (635/532) F Pixels B Pixels Circularity Sum of > Medians (635/532) Sum of Means (635/532) Log Ratio > (635/532) F635 Median - B635 F532 Median - B532 F635 Mean - > B635 F532 Mean - B532 F635 Total Intensity F532 Total > Intensity SNR 635 SNR 532 Flags Normalize Autoflag > > > >Can anybody help me out? We need to deal with GenePix data for our > experiment. > > > >Thanks in advance! > > > > > >Bin
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@gordon-smyth
Last seen 1 hour ago
WEHI, Melbourne, Australia
Er, why are you specifying sep="," for tab-delimited file? This file reads fine in all versions of limma using default settings. Gordon At 10:44 PM 17/10/2005, Ye, Bin wrote: >Hi, Gordon, > >Thank you for your reply! I feel it's more likely a limma problem, see my >code below: > >RG <- read.maimages("03_122589_0693_1_Cont_PMT400_r40_g10.gpr", >source="genepix", sep=",") >Error in "[.data.frame"(obj, , columns[[a]]) : > undefined columns selected > >The gpr file was generated by GenePix 6. I'm attaching the file, in case >you need it for debugging. > >BTW, my system is: R 2.1.1, limma 2.0.8 and limma 2.2.0 both used, both >gave the same error as above. > > >Bin > > >-----Original Message----- >From: Gordon Smyth [mailto:smyth at wehi.edu.au] >Sent: Sat 10/15/2005 6:20 AM >To: Ye, Bin >Cc: bioconductor at stat.math.ethz.ch; Saroj Mohapatra >Subject: Re: [BioC] Limma read GenePix version 6 file > >limma has been well tested with GenePix 6 and there should be no problem. >Specifying columns explicitly should not be required. If you believe >there's a limma error, please give the code you used so that we can debug. > >BTW, the columns that Saroj specified (all medians) can now be given as > > source="genepix.median" > >Gordon > > >Date: Fri, 14 Oct 2005 16:11:25 -0400 > >From: "Ye, Bin" <binye at="" med.wayne.edu=""> > >Subject: Re: [BioC] Limma read GenePix version 6 file > >To: "Saroj Mohapatra" <smohapat at="" vbi.vt.edu=""> > >Cc: bioconductor at stat.math.ethz.ch > > > >It works! Thank you, Saroj! > > > >Have a good weekend! > > > > > > > >Bin > > > > > >-----Original Message----- > >From: Saroj Mohapatra [mailto:smohapat at vbi.vt.edu] > >Sent: Fri 10/14/2005 3:19 PM > >To: Ye, Bin > >Cc: bioconductor at stat.math.ethz.ch > >Subject: Re: [BioC] Limma read GenePix version 6 file > > > >Bin: > > > >Which function are you using for reading the data? > > > >If you are using the limma function read.maimages, then the function > >provides some extra arguments that can force it to read specific > >columns. The help file says: > > > >---- > > > ?read.maimages() > > > >columns: list with fields 'Rf', 'Gf', 'Rb' and 'Gb' giving the column > > names to be used for red and green foreground and background > > or, in the case of Imagene data, a list with fields 'f' and > > 'b'. This argument is not usually specified by the user but, > > if it is, it over-rides 'source'. > >----- > > > >Something like this might work: > > > >myRG = read.maimages(myGenepix6Files, columns=list(Rf='F635 Median', > >Gf='F532 Median', Rb = 'B635 Median', Gb='B532 Median'), ...) > > > >Also, there are other arguments (other.columns) in the function for > >reading more columns from the file. > > > >Hope it helps. Best, > > > >Saroj > > > >Ye, Bin wrote: > > > > >Hi, > > > > > >When I try to read the GenePix output file from GenePix 6, I got an > > error as below: > > > > > >Error in "[.data.frame"(obj, , columns[[a]]) : > > > undefined columns selected > > > > > >I suspect GenePix 6 have some new columns different than previous version: > > > > > >Block Column Row Name ID X Y Dia. F635 > > Median F635 Mean F635 SD F635 CV B635 B635 Median B635 > > Mean B635 SD B635 CV % > B635+1SD % > B635+2SD F635 % > > Sat. F532 Median F532 Mean F532 SD F532 CV B532 B532 > > Median B532 Mean B532 SD B532 CV % > B532+1SD % > > > B532+2SD F532 % Sat. Ratio of Medians (635/532) Ratio of > > Means (635/532) Median of Ratios (635/532) Mean of Ratios > > (635/532) Ratios SD (635/532) Rgn Ratio (635/532) Rgn R2 > > (635/532) F Pixels B Pixels Circularity Sum of > > Medians (635/532) Sum of Means (635/532) Log Ratio > > (635/532) F635 Median - B635 F532 Median - B532 F635 Mean - > > B635 F532 Mean - B532 F635 Total Intensity F532 Total > > Intensity SNR 635 SNR 532 Flags Normalize Autoflag > > > > > >Can anybody help me out? We need to deal with GenePix data for our > > experiment. > > > > > >Thanks in advance! > > > > > > > > >Bin
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