Analysis of Differentially Expressed Genes using Microarray Technology
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@eleonora-lusito-5489
Last seen 9.6 years ago
Dear BioC users, I have a question regarding microarray data analysis (Human Affymetrix one color). My point is that I have just 1 sample TREATMENT and 1 sample REFERENCE. Neither technical replicates nor biological replicates are available. A statistical test to find differentially expressed genes between the two conditions seems to me impossible (even the simple t-test) due to the absence of replicates. People who asked me to do the analysis were interested only in finding the genes changing between the two conditions. In this conditions, in my opinion only the fold change is possible just to give a general view of the behavior of the genes. Any other suggestion about this issue? Thanks a lot E. -- Eleonora Lusito Computational Biology PhD student Molecular Medicine Program via Ripamonti 435, 20141 Milano, Italy Phone number: +390294375160 e-mail: eleonora.lusito at ieo.eu
Microarray Microarray • 1.2k views
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@richard-friedman-513
Last seen 9.6 years ago
Dear Eleonora, The results will not be reliable because you will not take the variability (both biological and technical) into account. One replicate each will only give you log Foldchanges, but no measure of the variability. Whether you go ahead or not is up to you but I try to avoid doing this whenever possible and I also tell the people that any such results come with a big question mark. with hope that this helps, Rich Richard A. Friedman, PhD Associate Research Scientist, Biomedical Informatics Shared Resource Herbert Irving Comprehensive Cancer Center (HICCC) Lecturer, Department of Biomedical Informatics (DBMI) Educational Coordinator, Center for Computational Biology and Bioinformatics (C2B2)/ National Center for Multiscale Analysis of Genomic Networks (MAGNet) Room 824 Irving Cancer Research Center Columbia University 1130 St. Nicholas Ave New York, NY 10032 (212)851-4765 (voice) friedman@cancercenter.columbia.edu http://cancercenter.columbia.edu/~friedman/ In memoriam, Ray Bradbury On Sep 10, 2012, at 2:38 PM, Eleonora Lusito wrote: > Dear BioC users, I have a question regarding microarray data analysis (Human > Affymetrix one color). My point is that I have just 1 sample TREATMENT and 1 > sample REFERENCE. Neither technical replicates nor biological replicates are > available. A statistical test to find differentially expressed genes between > the two conditions seems to me impossible (even the simple t-test) due to the > absence of replicates. People who asked me to do the analysis were interested > only in finding the genes changing between the two conditions. In this > conditions, in my opinion only the fold change is possible just to give a > general view of the behavior of the genes. Any other suggestion about this > issue? > > Thanks a lot > > E. > > -- > Eleonora Lusito > Computational Biology PhD student > Molecular Medicine Program > via Ripamonti 435, 20141 Milano, Italy > > Phone number: +390294375160 > e-mail: eleonora.lusito@ieo.eu > > _______________________________________________ > Bioconductor mailing list > Bioconductor@r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor [[alternative HTML version deleted]]
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Dear Dott. Friedman, I TOTALLY AGREE with you!! I stressed the people gave me the data to do at least 3 replicates per condition but samples are not available anymore, unfortunately.. Best E. Richard Friedman (friedman at cancercenter.columbia.edu) wrote: > > Dear Eleonora, > > The results will not be reliable because you will not > take the variability (both biological and technical) into account. > One replicate each will only give you log Foldchanges, but no > measure of the variability. Whether you go ahead or not > is up to you but I try to avoid doing this whenever possible > and I also tell the people that any such results come with > a big question mark. > > with hope that this helps, > Rich > Richard A. Friedman, PhD > Associate Research Scientist, > Biomedical Informatics Shared Resource > Herbert Irving Comprehensive Cancer Center (HICCC) > Lecturer, > Department of Biomedical Informatics (DBMI) > Educational Coordinator, > Center for Computational Biology and Bioinformatics (C2B2)/ > National Center for Multiscale Analysis of Genomic Networks (MAGNet) > Room 824 > Irving Cancer Research Center > Columbia University > 1130 St. Nicholas Ave > New York, NY 10032 > (212)851-4765 (voice) > friedman at cancercenter.columbia.edu > http://cancercenter.columbia.edu/~friedman/ > > In memoriam, Ray Bradbury > > On Sep 10, 2012, at 2:38 PM, Eleonora Lusito wrote: > > > Dear BioC users, I have a question regarding microarray data analysis (Human > > Affymetrix one color). My point is that I have just 1 sample TREATMENT and 1 > > sample REFERENCE. Neither technical replicates nor biological replicates are > > available. A statistical test to find differentially expressed genes between > > the two conditions seems to me impossible (even the simple t-test) due to the > > absence of replicates. People who asked me to do the analysis were interested > > only in finding the genes changing between the two conditions. In this > > conditions, in my opinion only the fold change is possible just to give a > > general view of the behavior of the genes. Any other suggestion about this > > issue? > > > > Thanks a lot > > > > E. > > > > -- > > Eleonora Lusito > > Computational Biology PhD student > > Molecular Medicine Program > > via Ripamonti 435, 20141 Milano, Italy > > > > Phone number: +390294375160 > > e-mail: eleonora.lusito at ieo.eu > > > > _______________________________________________ > > Bioconductor mailing list > > Bioconductor at r-project.org > > https://stat.ethz.ch/mailman/listinfo/bioconductor > > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor > > -- Eleonora Lusito Computational Biology PhD student Molecular Medicine Program via Ripamonti 435, 20141 Milano, Italy Phone number: +390294375160 e-mail: eleonora.lusito at ieo.eu
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@james-w-macdonald-5106
Last seen 1 hour ago
United States
Hi Eleonora, On 9/10/2012 2:38 PM, Eleonora Lusito wrote: > Dear BioC users, I have a question regarding microarray data analysis (Human > Affymetrix one color). My point is that I have just 1 sample TREATMENT and 1 > sample REFERENCE. Neither technical replicates nor biological replicates are > available. A statistical test to find differentially expressed genes between > the two conditions seems to me impossible (even the simple t-test) due to the > absence of replicates. People who asked me to do the analysis were interested > only in finding the genes changing between the two conditions. In this > conditions, in my opinion only the fold change is possible just to give a > general view of the behavior of the genes. Any other suggestion about this > issue? You are correct - all you can do is rank by fold change. Best, Jim > > Thanks a lot > > E. > -- James W. MacDonald, M.S. Biostatistician University of Washington Environmental and Occupational Health Sciences 4225 Roosevelt Way NE, # 100 Seattle WA 98105-6099
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@thomas-hampton-2820
Last seen 9.6 years ago
This is a great question. Obviously, with an N of 1, you are sorely limited in what you can say. In your consideration, you need to consider the ratio between your two samples, but you should also consider whether the ratio is based on a gene that is well-expressed in your system. Most genes are not very well expressed, and so many of your "largest" ratios will involve genes that are expressed at very low levels -- low enough so you might wonder whether the ratio is just pure noise. If I were you, I would look first at genes that are well expressed and show a large ratio, as well. You may also want to place emphasis on genes that are well understood and well annotated. At the end of the day, you can only make very preliminary statements of course, but you may see something that is worth following up on... Best Tom On Sep 10, 2012, at 2:38 PM, Eleonora Lusito wrote: > Dear BioC users, I have a question regarding microarray data analysis (Human > Affymetrix one color). My point is that I have just 1 sample TREATMENT and 1 > sample REFERENCE. Neither technical replicates nor biological replicates are > available. A statistical test to find differentially expressed genes between > the two conditions seems to me impossible (even the simple t-test) due to the > absence of replicates. People who asked me to do the analysis were interested > only in finding the genes changing between the two conditions. In this > conditions, in my opinion only the fold change is possible just to give a > general view of the behavior of the genes. Any other suggestion about this > issue? > > Thanks a lot > > E. > > -- > Eleonora Lusito > Computational Biology PhD student > Molecular Medicine Program > via Ripamonti 435, 20141 Milano, Italy > > Phone number: +390294375160 > e-mail: eleonora.lusito at ieo.eu > > _______________________________________________ > Bioconductor mailing list > Bioconductor at r-project.org > https://stat.ethz.ch/mailman/listinfo/bioconductor > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor >
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Hi Tom, thanks a lot for your suggestions! I'll ask them for a marker! Thanks again Best E. Thomas H. Hampton (Thomas.H.Hampton at dartmouth.edu) wrote: > > This is a great question. Obviously, with an N of 1, you are sorely limited in what > you can say. In your consideration, you need to consider the ratio between your > two samples, but you should also consider whether the ratio is based on a gene > that is well-expressed in your system. Most genes are not very well expressed, > and so many of your "largest" ratios will involve genes that are expressed at very > low levels -- low enough so you might wonder whether the ratio is just pure > noise. If I were you, I would look first at genes that are well expressed and show a large > ratio, as well. You may also want to place emphasis on genes that are well understood > and well annotated. At the end of the day, you can only make very preliminary statements > of course, but you may see something that is worth following up on... > > Best > > Tom > On Sep 10, 2012, at 2:38 PM, Eleonora Lusito wrote: > > > Dear BioC users, I have a question regarding microarray data analysis (Human > > Affymetrix one color). My point is that I have just 1 sample TREATMENT and 1 > > sample REFERENCE. Neither technical replicates nor biological replicates are > > available. A statistical test to find differentially expressed genes between > > the two conditions seems to me impossible (even the simple t-test) due to the > > absence of replicates. People who asked me to do the analysis were interested > > only in finding the genes changing between the two conditions. In this > > conditions, in my opinion only the fold change is possible just to give a > > general view of the behavior of the genes. Any other suggestion about this > > issue? > > > > Thanks a lot > > > > E. > > > > -- > > Eleonora Lusito > > Computational Biology PhD student > > Molecular Medicine Program > > via Ripamonti 435, 20141 Milano, Italy > > > > Phone number: +390294375160 > > e-mail: eleonora.lusito at ieo.eu > > > > _______________________________________________ > > Bioconductor mailing list > > Bioconductor at r-project.org > > https://stat.ethz.ch/mailman/listinfo/bioconductor > > Search the archives: http://news.gmane.org/gmane.science.biology.informatics.conductor > > > > > -- Eleonora Lusito Computational Biology PhD student Molecular Medicine Program via Ripamonti 435, 20141 Milano, Italy Phone number: +390294375160 e-mail: eleonora.lusito at ieo.eu
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