Newbie question regarding SAM analysis
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@ettinger-nicholas-1549
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Simon Lin ▴ 270
@simon-lin-1272
Last seen 10.0 years ago
Hi Nick: You are right. There is something weird about your data. Did you generate a simple pairwise plot (MvA or a simply scatter plot) of any pair of the arrays? From the SAM output, it seems that there is no differentially expressed genes. FDR are all close to 1, no matter what delta is, which means 100 percent identified (called) genes are false positive. There is only 18 called genes, even though delta is already very small, and the falsely identified one become larger than identified genes! Simon Message: 3 Date: Thu, 22 Dec 2005 11:51:55 -0600 From: "Ettinger, Nicholas" <nicholas-ettinger@uiowa.edu> Subject: [BioC] Newbie question regarding SAM analysis To: <bioconductor at="" stat.math.ethz.ch=""> Message-ID: <a4aa05ce92dacc43886d133db94a926e218cd063 at="" medicine-="" exch1.medicine.uiowa.edu=""> Content-Type: text/plain Hello all! This is my first post. Any help or suggestions would be greatly appreciated! I am trying to analyze 8 arrays (4 untreated, 4 treated; paired & alternating) with SAM. When I read the vignettes from 'siggenes' and looked at the sample diagrams, I was expecting to see my 'Called' column go from some number much closer to the number of probes on the hgu133probe2 Affy gene chip (something like 50,000 I think) down to zero. Why does it only start at 18? I am thoroughly confused by that. Thanks for any suggestions!! Happy Holidays to all!! ---Nick Ettinger University of Iowa Here is my code: TotalData <- ReadAffy() chipnumber <- length(sampleNames(TotalData)) chipnames <- sampleNames(TotalData) eset_rma <- rma(TotalData) K <- chipnumber/2 eset.cl <- rep(1:K, e = 2) * rep(c(-1, 1), K) eset.gnames <- geneNames(TotalData) sam.out <- sam(eset_rma, eset.cl, rand = 123, gene.names = eset.gnames) sam.out SAM Analysis for the Two-Class Paired Case Delta p0 False Called FDR 1 0.1 0.986 44.500 18 1 2 0.3 0.986 28.250 13 1 3 0.4 0.986 3.688 2 1 4 0.6 0.986 1.062 1 1 5 0.8 0.986 1.062 1 1 6 1.0 0.986 1.062 1 1 7 1.1 0.986 1.062 1 1 8 1.3 0.986 1.062 1 1 9 1.5 0.986 1.062 1 1 10 1.6 0.986 1.062 1 1 summary(sam.out) SAM Analysis for the Two-Class Paired Case s0 = 0.0646 (The 10 % quantile of the s values.) Number of permutations: 16 (complete permutation) MEAN number of falsely called genes is computed. Delta p0 False Called FDR cutlow cutup j2 j1 1 0.1 0.986 44.500 18 1 -4.503 6.912 16 54674 2 0.3 0.986 28.250 13 1 -5.004 6.912 11 54674 3 0.4 0.986 3.688 2 1 -7.102 Inf 2 54676 4 0.6 0.986 1.062 1 1 -8.690 Inf 1 54676 5 0.8 0.986 1.062 1 1 -8.690 Inf 1 54676 6 1.0 0.986 1.062 1 1 -8.690 Inf 1 54676 7 1.1 0.986 1.062 1 1 -8.690 Inf 1 54676 8 1.3 0.986 1.062 1 1 -8.690 Inf 1 54676 9 1.5 0.986 1.062 1 1 -8.690 Inf 1 54676 10 1.6 0.986 1.062 1 1 -8.690 Inf 1 5 4676
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