这是我的R包运行程序,我想以正常组为对照,您能帮我看下这样运行出来的是以正常为对照的吗?谢谢了。对于不一样的结果是取R包的还是DEcenter的?
> foldChange=1
> padj=0.05
>
> setwd("F:\\TCGA database RNAseq data difference analysis(none transference)\\none transference\\mRNA differenct analysis (none transference)") #设置工作目录
> library("edgeR")
载入需要的程辑包:limma
> rt=read.table("mRNA genesymbol.txt",sep="\t",header=T,check.names=F) #改成自己的文件名
> rt=as.matrix(rt)
> rownames(rt)=rt[,1]
> exp=rt[,2:ncol(rt)]
> dimnames=list(rownames(exp),colnames(exp))
> data=matrix(as.numeric(as.matrix(exp)),nrow=nrow(exp),dimnames=dimnames)
> data=avereps(data)
> data=data[rowMeans(data)>1,]
> group=c(rep("tumor",304),rep("normal",3)) #按照癌症和正常样品数目修改
> design <- model.matrix(~group)
> y <- DGEList(counts=data,group=group)
> y <- calcNormFactors(y)
> y <- estimateCommonDisp(y)
> y <- estimateTagwiseDisp(y)
> et <- exactTest(y,pair = c("tumor","normal"))
> topTags(et) ###这个是程序运行出来的
Comparison of groups: normal-tumor
logFC logCPM PValue FDR
TRPC4 6.251972 0.09996719 4.418785e-83 7.969279e-79
TCEAL6 8.622850 -2.39713054 1.545282e-82 1.393458e-78
CNN1 7.511701 5.42022391 3.949778e-77 2.374475e-73
PTGER3 7.278888 2.02023744 2.437630e-74 1.099067e-70
RBFOX3 6.366713 -0.96108111 1.533045e-72 5.529694e-69
CRHBP 6.399684 -1.72276640 1.128161e-70 3.391065e-67
LMOD1 6.453341 4.19172644 6.796867e-68 1.751164e-64
CLEC3B 5.694216 1.23873936 7.994085e-68 1.802166e-64
CSRP1 4.072072 8.03663267 8.245500e-66 1.652307e-62
HSPB7 6.321386 1.63355028 2.547360e-63 4.594163e-60
> ordered_tags <- topTags(et, n=100000)