Dear Dr. Sudheer Babu

You do not pool 12 values to obtain mean and SD. Instead, you should calculate the mean of three technical replicates to obtain mean of each biological replicate as this is considered a more robust approach compared to taking the mean of all technical replicates across all biological replicates, especially when analyzing variability.

If you only take the mean of all technical replicates across all biological replicates, you're essentially averaging out the biological variability, potentially masking significant differences between the biological groups. Please note that while technical replicates help to refine the measurements within each biological sample, biological replicates are crucial for understanding and interpreting the overall variability and differences between different experimental conditions or groups.

So, by taking the mean of 3 technical replicates for each of 4 biological replicates, it will allow you to assess both the biological variability between different samples and the technical variability within a single sample.

Best,

Malcolm Nobre Jochen Wilhelm

TIA.

Jochen Wilhelm