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siRNA with expensive gentamicin-containing medium

siRNA RNAi 184A1 viability

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#1 JDSBlueDevl



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Posted 24 March 2016 - 11:24 AM

I'm transfecting 184A1 immortalized mammary epithelial cells with siRNA using Lipofectamine RNAiMAX.  My problem is that the medium I use (MEGM) is expensive, so I can only order 1 at a time.  The medium I have right now contains gentamicin, per the instructions of the BulletKit.  When I talked to the tech at ThermoFisher, he expressed some concern regarding viability (I was more concerned about transfection efficiency).  However, when I looked at the cells 24 hrs after transfection, there did not seem to be a difference between the untransfected (that is, no treatment with RNAiMAX) and transfected wells.  I am going to wait another 24 hrs to check my knockdowns, as this is the first time I'm doing it with this protein and want to make sure I don't have any issues with a protein that could have a long half-life.  Is it possible I made it out of the "danger zone" and my cells will be just fine, or should I expect massive die-off tomorrow?

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