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Dual luciferase assay - proper controls?


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#1 greenhorn

greenhorn

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Posted 22 June 2011 - 12:07 PM

Hey everyone,

I am trying to perform a dual luciferase reporter assay in a cell line with promoters established from another cell line.
cDNA and protein of the gene are present in both cell lines.

Problem: In the new cell line, the luciferase activity for my promoters of interest is below pGL3, whereas TKpGL3 is as highly active as in the other cell line. Renilla activity was equal in all probes.

So I am looking for appropriate controls, to find out if this is sequence specific. Thought about beta-actin or GAPDH or any other gene naturally expressed in the cells...?

If someone has any ideas or suggestions on how to find out what's going on I'd be very grateful!

Thanks!!!




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