I'm trying to transfect CHO (and other cells later on) with a new construct. I've had success in the past with Lipofectamine on lung cancer cell lines and will probably try the same. I'm wondering about a reporter plasmid that I can use to cotransfect with my construct to monitor transfection efficiency. Should I use GFP or luciferase or something else? Also, where is the best place to get these plasmids? Thanks again.
Transfecting GFP plasmid in a cotransfection with your main one is a good positive control (https://altogen.com/product/gfp-expressing-plasmid-dna-25-ug/). However it does not necessarily mean that your main plasmid is also being expressed. Including GFP in your main plasmid does guarantee that GFP expression correlates with vector expression, especially if GFP is downstream from the target gene.
You could co-transfect with GFP/luciferase plasmids (or any reporter plasmid for that matter). GFP would be the easiest as you can check transfection efficiency (# of cells that were transfected out of a certain # in your visual field) using a fluorescent microscope. If you use flow cytometry, you could get a a more complete picture (frequencies among all your cells). Luciferase would give you a vague measure of transfection but it won't give you the frequencies of cells that were transfected.
In any case, as long as you use co-transfection, the fact that your cells express the reporter plasmid does not guarantee that they took up your plasmid of interest. The best thing to do would be to clone in GFP/luciferase into your construct of interest. This way, any cell that is reporter+ will have to express your gene of interest.
The most comprehensive source would be Addgene.
https://www.addgene.org/
I agree with Julie, co-transfecting a GFP expressing plasmid would be the more convenient ; when it comes to transfection rate evaluation, visual inspection by fluorescence microscopy stays the best way: easy, cost effective and rather accurate.
you could also take a look at this:
Article A simple, rapid method for evaluation of transfection effici...
Transfecting GFP plasmid in a cotransfection with your main one is a good positive control (https://altogen.com/product/gfp-expressing-plasmid-dna-25-ug/). However it does not necessarily mean that your main plasmid is also being expressed. Including GFP in your main plasmid does guarantee that GFP expression correlates with vector expression, especially if GFP is downstream from the target gene.
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