Created
August 14, 2021 20:14
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library(DESeq2) | |
#/ from https://raw.githubusercontent.com/shangguandong1996/picture_link/main/WFX_count_Rmatrix.txt | |
data <- read.delim("WFX_count_Rmatrix.txt", row.names = "Geneid") | |
get_res <- function(dds){ | |
dds <- DESeq(dds) | |
res <- results(dds, name=resultsNames(dds)[2]) | |
lfcShrink(dds, coef=resultsNames(dds)[2], res=res, type="ashr") | |
} | |
#/ original data: | |
res1 <- | |
get_res( | |
DESeqDataSetFromMatrix( | |
countData = ceiling(data), | |
colData = data.frame(condition=factor(rep(c("A", "B"), each=2))), | |
design = ~condition) | |
) | |
#/ prefiltered: | |
res2 <- | |
get_res( | |
DESeqDataSetFromMatrix( | |
countData = ceiling(data)[rowSums(data > 10 ) >= 2,], | |
colData = data.frame(condition=factor(rep(c("A", "B"), each=2))), | |
design = ~condition) | |
) | |
#/ the same with simply copy/pasting the samples to make up more replicates, | |
#/ now with 20 samples per group: | |
res3 <- get_res( | |
DESeqDataSetFromMatrix( | |
countData = ceiling(do.call(cbind, lapply(1:10, function(x) data))), | |
colData = data.frame(condition=factor(rep(rep(c("A", "B"), each=2), 10))), | |
design = ~condition) | |
) | |
#/ MA-plots are essentially the same towards odd shape, | |
#/ color is turned off here to not look messy | |
pdf("maplots.pdf") | |
par(mfrow=c(2,2)) | |
plotMA(res1, alpha=0, ylim=c(-10,10), main="original") | |
plotMA(res2, alpha=0, ylim=c(-10,10), main="prefiltered") | |
plotMA(res3, alpha=0, ylim=c(-10,10), main="20 samples per group") | |
dev.off() | |
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