Summarizing biological replicates for RNASeq

[BioInfoBee]

Hello All,
I have three biological replicates for each single tissue sample of RNASeq data? Assuming that these each biological replicates are normalized (RPKM, TPM), is there any other statistical analysis that needs to be taken into account before deciding on taking average (mean), median or other approaches?

For example,

S1.1 S1.2 S1.3
Gene1 0 0 9
Gene2 3 3 3
Gene3 2 6 1

Kindly suggest!

[GenomicSeq]

BioInfoBee Yes, before taking the average or median, you should conduct a variance or dispersion analysis (like calculating the coefficient of variation) across the biological replicates to ensure that they are consistent with each other. If there's high variability between replicates, it might not be appropriate to simply average them.

[BioInfoBee]

GenomicSeq Thank you, for your feedback. But, what if there exhibit a relatively high degree of variability between biological (3) replicates. For example, CV for above example;:

S1.1 S1.2 S1.3 CV
Gene1 0 0 9 141.42%
Gene2 3 3 3 0.0%
Gene3 2 6 1 72.01%

In such case what could be the alternative taking an average (mean) of biological replicates?

Regards,.

[BasesBreaker]

In this case, it's probably more appropriate to use the median of the replicates instead of the mean. I'm not the greatest with this type of analysis, but the median is a robust statistic that is less sensitive to outliers and will provide a more representative value in the presence of such variability.