Just wondering if it is better to use a merged transcriptome of the samples I have or should I use the human reference annotation? I intend to do differential expression so I was planning on using RSEM to calculate counts. I know that the merged transcriptome might result in novel transcripts but I was wondering if it still would be a fair analysis to just use the human reference annotation?
2 answers
Depends if you want to check only known genes/transcripts or you want to consider the presence of new unknown transcripts/isoforms. From the research point of view you want to do both, then compare the differences and see if there are any advantages from one annotation to the other.
For human, the reference annotation should be better than what you could work out experimentally. So if you aren't looking for novel transcripts, just use the annotation that already exists.
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