How many minimum biological replicates are required to compare the transcriptome of two conditions?
Hi Narendra,
you mean 1 sample vs 1 sample? If so, the answer is no, because you need biological replicates to generate a statistical analysis and RNA-Seq is no exception.
The number of replicates required to build a solid RNA-Seq analysis depends on many factors, among them the heterogeneity of the replicates. As a rule of thumb you need at least 3; 4-5 is better.
Regards,
Rocco
Using one sample for RNA-seq analysis would be a big waste of money, efforts and resources.
There is a publication about this topic
Article How many biological replicates are needed in an RNA-seq expe...
Technical replicates can be used if you don't have biological replicates. Or if there is a budget issue, better to go for detailed qPCR analysis rather than investing in RNAseq.
Dear Rocco Piazza,
Many thanks for answering my query.
Yes, I mean 1 sample vs 1 sample RNA-Seq analysis.
Regards,
Narendra
Dear Abhijeet Singh,
Many thanks for answering my query.
Yes, Schurch et al. 2016 is an excellent paper to decide the number of biological replicates for RNA-Seq analysis.
Regards,
Narendra
In my opinion, it also depends on how depth you performed your sequencing . If there is low coverage, you need more samples, such as more than 3 replicates, etc. If there is a high coverage, a minimum 3 replicates is enough , which just likes many other biological bench work. As to the single sample from each condition, there will be a limited use of the date as a reference to show the presence of the gene expression (absence, not sure), for example, but in quantities it shoud be verified by real-time PCR.
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