For example, in some experiments I transfect cells with plasmid DNA to produce retroviruses pseudotyped with other viral envelope proteins. I am using X-tremeGENE HP, which does not require the medium to be changed following transfection. Of course, it is much easier to not have to change the media, so I didn't. 48hrs post-transfection, I filtered this virus-containing supernatant through a 0.45um filter to be subsequently used for infecting naive cells.
Is there a chance these new naive cells could be transfected with reagent: DNA complexes remaining in the medium? Or will such complexes degrade over 48 hrs in a CO2 incubator?
Does anyone have any experience or ideas with this?
If you want to follow infections specifically, I would highly recommend you to change media about 6-12 hrs post transfection. If you dont change the media, and continue with your retroviral transduction, you will definitely get transfection from the remaining DNA complexes, however little you have. Even if there are no transfections occuring, you can never be certain that you are seeing bonafide infections. In my case, I observed between 5-20% of transfection of the "transduced" cells, if I did not change the media after the initial transfection.
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