I have a question on averaging biological replicates together. the code below plots data for each sample. When and how do I combine my biological replicates for plotting? I would like to combine over the CONDITION column and have tried a man ways. Thought you might have an answer.
colData(rld)
DataFrame with 6 rows and 10 columns
sampleName fileName LINE EXPOSURE CONDITION TISSUE REP
<factor> <factor> <factor> <factor> <factor> <factor> <integer>
A1H_Acute A1H_Acute A1H_Acute CSB Acute Cocaine H 1
A2H_Acute A2H_Acute A2H_Acute CSB Acute Cocaine H 2
A3H_Acute A3H_Acute A3H_Acute CSB Acute Cocaine H 3
B1H_Acute B1H_Acute B1H_Acute CSB Acute Sucrose_C H 1
B2H_Acute B2H_Acute B2H_Acute CSB Acute Sucrose_C H 2
B3H_Acute B3H_Acute B3H_Acute CSB Acute Sucrose_C H 3
SEX individual sizeFactor
<factor> <factor> <numeric>
A1H_Acute M AM 1.23895646591537
A2H_Acute M AM 0.709636373005609
A3H_Acute M AM 1.39159832544129
B1H_Acute M BM 0.738832280319489
B2H_Acute M BM 0.908432365721923
B3H_Acute M BM 1.24898796150053
dds <- DESeqDataSetFromMatrix(countData = AcuteCountsMheadCO, colData = AcuteSampleTable1MheadCO, design = ~ CONDITION )
myTest<-DESeq(dds)
rld <- rlog(myTest, blind=F)
select <- order(rowMeans(counts(myTest,normalized=TRUE)),
decreasing=TRUE)[1:20]
df <- as.data.frame(colData(myTest)[,c("CONDITION","TISSUE")])
pheatmap(assay(rld)[select,], cluster_rows=FALSE, show_rownames=FALSE,
cluster_cols=FALSE, annotation_col=df)
Hi James,
I realize my code is not correct in the aspect of plotting the most significant DEGenes. I am very new to DEseq2 and making heat maps. In the end I want to generate a heat map for the "pulled" samples on only the most significant genes. In this case it was 104 genes.