I would like to check the transcript copy of the GFP-tagged chimeric protein that I have introduced into the animal via viral vector via RT-qPCR. Although
I can see the GFP in the histology of the sectioned tissue, I am looking for another method to quantify the expression of this chimeric protein.
The tissue of interest is too small to yield sufficient amount of protein for a Western blot and still have enough for other assays. Would it possible to reliably detect eGFP expression level via qRT-PCR assay?
Yes it is possible to detect EGFP in expressing cells/tissues by RT-qPCR. I did this for single cells in the paper linked here.
You will need to design primers for the specific sequence of your EGFP. Do you have any information on your particular variant? (like an original plasmid?) Designing the primers is fairly trivial and I would be happy to help.
Article A convenient, optimized pipeline for isolation, fluorescence...
Yes it is possible to detect EGFP in expressing cells/tissues by RT-qPCR. I did this for single cells in the paper linked here.
You will need to design primers for the specific sequence of your EGFP. Do you have any information on your particular variant? (like an original plasmid?) Designing the primers is fairly trivial and I would be happy to help.
Article A convenient, optimized pipeline for isolation, fluorescence...
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