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How long after transfection or dox induction of stable cell line before performi

Transient Transfection Cell Cycle Dox induction

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#1 5280

5280

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Posted 15 February 2015 - 02:55 PM

I was wondering what a good time point is for performing cell cycle analysis after transiently transfecting human cells or dox induction of a stable cell line? This is in reference to overexpression of a transcription factor that regulates genes involved in cell cycle. Would 72hrs be sufficient to see alterations in cell cycle?

 

Trying to determine the optimal endpoint from overexpression until cell cycle harvest from steps 1-3: 72hrs?

 

1. Overexpress gene of interest/functional protein

2. Transactivation of targets genes involved in cell cycle regulation

3. Cell cycle analysis for alterations.

 

Just now sure how many hours are needed to see this effect?

 

Thanks!



#2 bob1

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Excellent

Posted 15 February 2015 - 05:50 PM

Probably depends on the gene and the promoter running it, but in general for transfections 24-48h should be enough, but I don't know for inducible genes, but I would say 48h or more.







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