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## In this example, the data is in a matrix called
## data.matrix
## columns are individual samples (i.e. cells)
## rows are measurements taken for all the samples (i.e. genes)
## Just for the sake of the example, here's some made up data...
data.matrix <- matrix(nrow=100, ncol=10)
colnames(data.matrix) <- c(
paste("wt", 1:5, sep=""),
paste("ko", 1:5, sep=""))
rownames(data.matrix) <- paste("gene", 1:100, sep="")
for (i in 1:100) {
wt.values <- rpois(5, lambda=sample(x=10:1000, size=1))
ko.values <- rpois(5, lambda=sample(x=10:1000, size=1))
data.matrix[i,] <- c(wt.values, ko.values)
}
head(data.matrix)
dim(data.matrix)
pca <- prcomp(t(data.matrix), scale=TRUE)
## plot pc1 and pc2
plot(pca$x[,1], pca$x[,2])
## make a scree plot
pca.var <- pca$sdev^2
pca.var.per <- round(pca.var/sum(pca.var)*100, 1)
barplot(pca.var.per, main="Scree Plot", xlab="Principal Component", ylab="Percent Variation")
## now make a fancy looking plot that shows the PCs and the variation:
library(ggplot2)
pca.data <- data.frame(Sample=rownames(pca$x),
X=pca$x[,1],
Y=pca$x[,2])
pca.data
ggplot(data=pca.data, aes(x=X, y=Y, label=Sample)) +
geom_text() +
xlab(paste("PC1 - ", pca.var.per[1], "%", sep="")) +
ylab(paste("PC2 - ", pca.var.per[2], "%", sep="")) +
theme_bw() +
ggtitle("My PCA Graph")
## get the name of the top 10 measurements (genes) that contribute
## most to pc1.
loading_scores <- pca$rotation[,1]
gene_scores <- abs(loading_scores) ## get the magnitudes
gene_score_ranked <- sort(gene_scores, decreasing=TRUE)
top_10_genes <- names(gene_score_ranked[1:10])
top_10_genes ## show the names of the top 10 genes
pca$rotation[top_10_genes,1] ## show the scores (and +/- sign)
#######
##
## NOTE: Everything that follow is just bonus stuff.
## It simply demonstrates how to get the same
## results using "svd()" (Singular Value Decomposition) or using "eigen()"
## (Eigen Decomposition).
##
#######
############################################
##
## Now let's do the same thing with svd()
##
## svd() returns three things
## v = the "rotation" that prcomp() returns, this is a matrix of eigenvectors
## in other words, a matrix of loading scores
## u = this is similar to the "x" that prcomp() returns. In other words,
## sum(the rotation * the original data), but compressed to the unit vector
## You can spread it out by multiplying by "d"
## d = this is similar to the "sdev" value that prcomp() returns (and thus
## related to the eigen values), but not
## scaled by sample size in an unbiased way (ie. 1/(n-1)).
## For prcomp(), sdev = sqrt(var) = sqrt(ss(fit)/(n-1))
## For svd(), d = sqrt(ss(fit))
##
############################################
svd.stuff <- svd(scale(t(data.matrix), center=TRUE))
## calculate the PCs
svd.data <- data.frame(Sample=colnames(data.matrix),
X=(svd.stuff$u[,1] * svd.stuff$d[1]),
Y=(svd.stuff$u[,2] * svd.stuff$d[2]))
svd.data
## alternatively, we could compute the PCs with the eigen vectors and the
## original data
svd.pcs <- t(t(svd.stuff$v) %*% t(scale(t(data.matrix), center=TRUE)))
svd.pcs[,1:2] ## the first to principal components
svd.df <- ncol(data.matrix) - 1
svd.var <- svd.stuff$d^2 / svd.df
svd.var.per <- round(svd.var/sum(svd.var)*100, 1)
ggplot(data=svd.data, aes(x=X, y=Y, label=Sample)) +
geom_text() +
xlab(paste("PC1 - ", svd.var.per[1], "%", sep="")) +
ylab(paste("PC2 - ", svd.var.per[2], "%", sep="")) +
theme_bw() +
ggtitle("svd(scale(t(data.matrix), center=TRUE)")
############################################
##
## Now let's do the same thing with eigen()
##
## eigen() returns two things...
## vectors = eigen vectors (vectors of loading scores)
## NOTE: pcs = sum(loading scores * values for sample)
## values = eigen values
##
############################################
cov.mat <- cov(scale(t(data.matrix), center=TRUE))
dim(cov.mat)
## since the covariance matrix is symmetric, we can tell eigen() to just
## work on the lower triangle with "symmetric=TRUE"
eigen.stuff <- eigen(cov.mat, symmetric=TRUE)
dim(eigen.stuff$vectors)
head(eigen.stuff$vectors[,1:2])
eigen.pcs <- t(t(eigen.stuff$vectors) %*% t(scale(t(data.matrix), center=TRUE)))
eigen.pcs[,1:2]
eigen.data <- data.frame(Sample=rownames(eigen.pcs),
X=(-1 * eigen.pcs[,1]), ## eigen() flips the X-axis in this case, so we flip it back
Y=eigen.pcs[,2]) ## X axis will be PC1, Y axis will be PC2
eigen.data
eigen.var.per <- round(eigen.stuff$values/sum(eigen.stuff$values)*100, 1)
ggplot(data=eigen.data, aes(x=X, y=Y, label=Sample)) +
geom_text() +
xlab(paste("PC1 - ", eigen.var.per[1], "%", sep="")) +
ylab(paste("PC2 - ", eigen.var.per[2], "%", sep="")) +
theme_bw() +
ggtitle("eigen on cov(t(data.matrix))")
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