Dear all
I am plaing to do dual luciferase reporter assay. I have transfected my cell with reporter plasmid and my gene of interest. My cells were around 80% at the time of trsnfecrion but now become 100% just after 6 hrs. So can I spilit my cell into two dishes because I still need to keep for 20hrs after changing medium. If i dnt spilit them they will become condluence. So, my question is does the spilit will effect my totall plasmid does. I used different dose of my desired gene like 300ng, 600ng, 900ng. Thanks
If you aren't planning on transfecting the population further (for sake of homogeneity) then you should be fine - there's nothing inherently wrong with passaging overly confluent cells. If you are planning on doing multiple transfections, like for the generation of stable cell lines (see https://altogen.com/services/generation-stably-expressing-cell-lines-28-days/ ) then you need to be clear about what kind of population you are seeking, and how your assay results could be affected.
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