I would like to localize a GFP reporter plasmid in mammalian cells by using DNA FISH. Can anyone recommend a probe sequence targeting GFP DNA?
Many thanks in advance.
Inci
Thank you Timothy for the answer.
The plasmid is about 8 Kb and will be delivered into the cell by pseudovirus particles. I have already tried labeling the reporter plasmid/pseudogenome with EdU and BrdU (in case you are referring to these kits), however these methods are not compatible with what I want to do: To look at the colocalization of the pseudogenome with a GFP-fused telomeric protein (unfortunately, immunofluorescent staining is not working properly, and EdU click iT reaction is not compatible with GFP fluorescence, and the EtOH/glycine fixation does not give good resolution of the telomeric protein). Therefore, I was thinking to use FISH with a telomere probe and a probe against the pseudogenome. What are things to consider, when designing FISH probes? Are there online tools helping to find good probes against a specific sequence? Would it be better to use several probes targeting different parts of the plasmid? This way one could also increase the signal?!
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