Is there an alternative to normalization for single-cell analysis?
The good thing is that you know that you’re measuring just one cell, so the main problem when you do these kinds of experiments is how to know that you’re getting all the mRNA out of the cell or not. That is very hard to determine. I have not done these experiments, but one way would be to inject a known amount of an alien mRNA molecule, and then you do this in a lot of cells and see if you get out the same amount when you lyse them … but that is something that no one has done. That is a little bit tricky to do. The normal way is actually that you know that you have one cell. With traditional [gene expression profiling], you maybe take 10,000 cells and then you extract [the DNA], but then you lose a lot of material so you can’t really compare it to 10,000 cells any more, so therefore you go on with these reference genes [and normalization]. So therefore the value you actually get out is copy numbers for one specific cell. So either you can deliver the data as the absolute number of cDN
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