PCA plot in DESeq2
1
0
Entering edit mode
pkachroo ▴ 10
@pkachroo-11576
Last seen 4.1 years ago

Hi,

For RNASeq analysis, I am generating a PCA plot for various strains with three biological replicates each. When I make the PCA plot , I get a symbol on the plot for every replicate. For a large dataset, I was wondering if there is a way to have a single symbol (average of three biological replicates) be represented on the plot, instead of all three replicates. 

In DESeq2 package I use:

 library(ggplot2)
 data <- plotPCA(rld, intgroup=c("clade", "strain"), returnData=TRUE)
 percentVar <- round(100 * attr(data, "percentVar"))
 ggplot(data, aes(PC1, PC2, color=strain, shape=clade)) +
  geom_point(size=3) +
  xlab(paste0("PC1: ",percentVar[1],"% variance")) +
  ylab(paste0("PC2: ",percentVar[2],"% variance")) +
  coord_fixed()

Thanks,

Priyanka 

 

deseq2 pca plot • 4.5k views
ADD COMMENT
2
Entering edit mode
@mikelove
Last seen 1 day ago
United States

I've received this question before on the support site, and my answer is that I really don't understand the point of a PCA plot in which you can't see how the samples within a group spread out. I suppose you can compare the distances between 3 or more conditions, but those distances relative to the biological variance are what I'm most interested in seeing in a PCA plot.

If you really want to make this plot despite these shortcoming I've mentioned, you can compute the row-wise average of the transformed values for each condition and make a PCA plot of just the means. The rowMeans() function can be used to for the means of a subset of the data, and cbind() can be used to bind the columns of means from the different groups together.

ADD COMMENT
0
Entering edit mode

Thanks Micheal. I complete agree with your reasoning. However, I have 30 samples in triplicates and visualizing the relationship between samples become difficult due to multiple data points. I intend to make both PCA plots, with individual replicates (to see spread within samples) and with average of replicates (spread between samples). 

ADD REPLY

Login before adding your answer.

Traffic: 722 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6