Hi guys,
When at the end of the analysis I export the results with this code:
write.csv(as.data.frame(resOrdered),
file="condition_treated_results.csv")
I have a file containing a list of genes with Log2FC and p-value. let say I have two conditions treated and untreated (6 and 2 ), are the values of gene expression of two replicates (untreated) normalized and compared with the values of the 6 replicates (treated) ? I mean are this taken as a single condition isn't it?
What if I have more conditions? let say 4 different conditions, to create that table 3 will be considered as a unique and one as reference condition?
Thank you
also, to make a heatmap where all the biological replicates cluster together I could take the top 1000 gene that are similar among replicates and different with the reference level but I don't know if that is right and/or DESeq2 can do that, what do you guys think?
Thanks
Thank you, now the first part is clear enough,
For the heatmap,I tried this code as you say but I have this result,
pheatmap(heatmap.genes) Error in hclust(d, method = method) : must have n >= 2 objects to cluster
I think is missing the information of the samples name which in my previous command for heatmao was rld,
Can you tell me how to fix it?
I used this to show the heatmap
library("pheatmap") pheatmap(heatmap.genes)
thank you
Looks like you’re having some issues going from the transformer data to the heatmap. I’d recommend go one step at a time and make sure you have what you are expecting at each step. Eg heatmap.genes is not data, it’s a set of numeric indices. You should get in the habit of looking at the various objects to see what they represent, using eg head() and dim(), etc.