Limited number of independent clones - how many 'independent' biological replicates?

[Alex234]

Hello, I am comparing transcription in KO and WT ES cell lines and am basically worried about pseudoreplication.

If a single cell line is split into two, and then they undergo many passages separated from each other, would that qualify as "having undergone independent growth"?

Would they then be considered independent biological replicates?


Many thanks

Alex

[dpryan]

Are these standard ES cell lines are did you make them yourself? The general question that you have to ask yourself regarding deciding what constitutes a proper replicate is, "If someone in on the other side of the world were to try to reproduce this, what sort of things would be different?" Your independent biological replicates should then be designed to reproduce this. If these are ES cells that you can get from ATCC or another source, then the situation you described is probably sufficient. If you made these ES cells yourself, then you'd want to use cells generated from multiple animals/people/whatever.