Hello. I am doing reporter assay experiments using the plasmid pGL4.10. I transfect with the empty, promoterless plasmid pGL4.10 and with PGL4.10 with a promoter of my interest. Moreover, I'm using the dual reporter assay system, so I always cotransfect with another plasmid with the Renilla luciferase gene.
I also infect with adenoviruses expressing a transcription factor. I will try to explain myself with a sample data:
-> Adenovirus CTL
PGL4.10-empty : Firefly/Renilla = 4,42 (A)
PGL4.10+promoter_genofinterest: Firefly/Renilla = 126,47 (B)
->Adenovirus transcription factor
PGL4.10-empty : Firefly/Renilla : Firefly/Renilla = 35,37 (C)
PGL4.10+promoter_genofinterest: Firefly/Renilla = 1737,19 (D)
I don't know how to manage the data correctly. I think that (D-C) / (B-A) maybe it's a correct way, but I'm not sure. If anyone has experience doing this kind of calculations I would be very grateful if you can advise me.
in my opinion if you want to calculate and presented the data of reporter gene assay (relative luciferase activity) , so it should be :
Control : A/B
Transcription group : C/D
than make bar diagram from the data and calculated how much the fold activation of transcription factor compared to control.
Thanks. I think so too but, in my case, the data is very difficult to interpret because of the great background (without inducing any transcription factor there are a lot of signal too).
Moreover, I don't know what statistical system is better for this approach. Multiple t-tests, maybe?
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