Hi all,
My name is Amy, I am a masters student in Bioinformatics at North Carolina
State University. I am working on a project and I am trying to use the lumi
R package for microarray data analysis. I have shown the sample code here
and have questions about modifying the sample code for my own data.
lumi package in R, example.lumi, the sample data has 8000 features and 4
samples
I have highlighted the code I have questions on in red, my data has 4
different types of samples, each repeated 6 times, so a total of 24 samples
and about 48,000 rows. how should I identify my sampleType in my case? also
what does colnames(design) <- c('100:0', '95:5-100:0') do, which columns
exactly does it take into consideration? Thanks!
so the sample code i'm trying to follow is below:
###################################################
### code chunk number 30: filtering
###################################################
presentCount <- detectionCall(example.lumi)
selDataMatrix <- dataMatrix[presentCount > 0,]
probeList <- rownames(selDataMatrix)
###################################################
### code chunk number 31: Identify differentially expressed genes
###################################################
## Specify the sample type
sampleType <- c('100:0', '95:5', '100:0', '95:5')
if (require(limma)) {
## compare '95:5' and '100:0'
design <- model.matrix(~ factor(sampleType))
colnames(design) <- c('100:0', '95:5-100:0')
fit <- lmFit(selDataMatrix, design)
fit <- eBayes(fit)
## Add gene symbols to gene properties
if (require(lumiHumanAll.db) & require(annotate)) {
geneSymbol <- getSYMBOL(probeList, 'lumiHumanAll.db')
geneName <- sapply(lookUp(probeList, 'lumiHumanAll.db',
'GENENAME'), function(x) x[1])
fit$genes <- data.frame(ID= probeList,
geneSymbol=geneSymbol, geneName=geneName, stringsAsFactors=FALSE)
}
## print the top 10 genes
print(topTable(fit, coef='95:5-100:0', adjust='fdr',
number=10))
## get significant gene list with FDR adjusted p.values
less than 0.01
p.adj <-
p.adjust(fit$p.value[,2])
sigGene.adj <- probeList[ p.adj < 0.01]
## without FDR adjustment
sigGene <- probeList[ fit$p.value[,2] < 0.01]
}
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