A puzzle about the results of differential genes
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llkxiaolan ▴ 10
@llkxiaolan-13767
Last seen 6.0 years ago

I use the limma package to analyze the data, extract the genetic differences, but in the list of results, adj.p.value are basically the same, which is why, with the same code, analysis of other data, it did not encounter this problem, it is the problem of data. 

Here are the results of  data analyzed using the code :

  logFC AveExpr t P.Value adj.P.Val B
1449938_at 0.913685 9.095604 7.035732 0.000213 0.999936 -2.24783
1424967_x_at 1.136483 5.485582 6.265933 0.000432 0.999936 -2.37058
1460719_a_at 1.267538 8.413155 5.994509 0.000563 0.999936 -2.42196
1449937_at 0.958864 8.878113 5.92327 0.000605 0.999936 -2.43625
1418726_a_at 0.890958 4.738728 5.710449 0.00075 0.999936 -2.4811
1427004_at 0.90456 5.830647 5.64532 0.000802 0.999936 -2.4955
1419758_at 1.167344 7.691656 5.599396 0.000841 0.999936 -2.50585
1418086_at 0.988556 6.526427 5.59649 0.000844 0.999936 -2.50651
1422411_s_at 0.96971 7.66573 5.377632 0.001063 0.999936 -2.55825

 Here is the code I use:

 library(affy)
AffyRawData<-ReadAffy(widget=TRUE)
data.class(AffyRawData)
par(mfrow=c(2,3))
image(AffyRawData[,c(1,2,3,4,5,6)])
library(simpleaffy)
Data.qc<-qc(AffyRawData)
plot(Data.qc)
library(RColorBrewer)
library(affyPLM)
Pset<-fitPLM(AffyRawData)
colors<-brewer.pal(12,"Set3")
Mbox(Pset,ylim=c(-1,1),col=colors,main="RLE",las=3)
boxplot(Pset,ylim=c(0.95,1.22),col=colors,main="NUSE",las=3)
data.deg<-AffyRNAdeg(AffyRawData)
plotAffyRNAdeg(data.deg,col=colors)
legend("topleft",rownames(pData(AffyRawData)),col=colors,lwd=1,inset=0.05,cex=0.5)
AffyRawDatarma<-rma(AffyRawData)
boxplot(AffyRawDatarma,col=colors,las=3,main="RMA")
library(limma)
eset<-exprs(AffyRawDatarma)
design <- model.matrix(~ 0+factor(c(1,1,1,2,2,2)))
colnames(design) <- c("group1", "group2")
contrast.matrix <- makeContrasts(contrasts="group2-group1",levels=design)
design
fit <- lmFit(eset, design)
fit1<- contrasts.fit(fit, contrast.matrix)
fit2 <- eBayes(fit1)
dif<-topTable(fit2,coef="group2-group1",n=nrow(fit2),lfc=log2(1.5))
dif<-dif[dif[,"P.Value"]<0.05,]
head(dif)

Is there anyone who can help me explain this problem? Thank you very much 

                 

 

limma • 979 views
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@gordon-smyth
Last seen 1 hour ago
WEHI, Melbourne, Australia

This isn't a problem -- it just means there are no significantly differentially expressed genes.

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Entering edit mode

Sorry, I don't understand. You mean the difference is very small, right? I can't use adj.p.value as a filter condition. Should I use P.Valve? Thanks very much 

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Entering edit mode
llkxiaolan ▴ 10
@llkxiaolan-13767
Last seen 6.0 years ago

Sorry, I don't understand. You mean the difference is very small, right? I can't use adj.p.value as a filter condition. Should I use P.Valve? Thanks very much 

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