Hi all,
I am working on transfection experiments involving miRNA in mammalian cell lines and I have a doubt here. After optimizing my experiment, I am able to achieve good transfection efficieny by measuring the luciferase/fluorescence signal post-transfection.
I would like to seek for your suggestion on whether I should always check for transfection efficiency before I start any cellular assays? It is because to check the transfection efficiency, I will need to co-transfect my cells with the miRNA and the plasmid DNA containing the luciferase/fluorescence reporter system. If I can skip the plasmid DNA transfection, then I can skip the co-transfection and just focus on my miRNA transfection.
Hope to hear for any fruitful recommendation by you all.
Thank you all in advance!
Always include co-transfectrion with a reference reporter construct under transient transfection conditions. You need this to monitor the transfection efficiency (which can be extremely variable between the experiments).
In my opinion, you can skip the co-transfection with your reporter plasmid. You can just transfect the cells with miRNA and confirm the transfection by qPCR of transfected miRNA. So you can identify that transfection is working and miRNA level is increasing or decreasing whatever is required.
It is better to include always co-transfection but if you are confident enough (at least 3 times you got same results after co-transfection) and then if there has financial limitation, you can skip it.
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