I am doing a knockdown test in 96 well plates and then I need to extract RNA and protein from my cells. I'm not sure that the number of cells in 96 well plates is enough to these extractions. Would you please let me know how I can design my experiment?
Thanks
You can get enough RNA from cells in a 96 well format to do qPCR, but sufficient protein will depend on your readout. For western blot, you likely won't have enough protein to visualize GAPDH (using most common readout methods) but if you want to do cell staining and use flow-cytometry you can definitely analyze those cells' proteins.
Thanks.
Unfortunately I want to do western blot!
How many days can I keep the cells after siRNA knowdown?
If I can split the cells after 3 days, the number of cells might be enough for qPCR and western blot. Yes?
In our hands, siRNAs typically had the best effect 48 or 72 hrs post transfection, you may want to optimize this to see how your siRNA performs. We used 6-well dished for siRNA experiments and got enough sample to analyze both RNA and protein by western blot. Good luck!
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