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Co-transfection of luciferase reporter plasmid and constitutively expressed beta


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#1 WYF

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Posted 14 May 2010 - 01:40 AM

Hi all,

I am about to try transfecting my cells with 2 plasmids: First plasmid is the luciferase reporter plasmid that report on the activity of my drug treatment, and the second will be the constitutively expressed beta-gal plasmid to check for transfection efficiency.

I would like to know if anyone can suggest a good starting point with respect to the ratio of these 2 plasmids? I was thinking of luciferase:beta-gal to be 2:1, with the logic that if 1 part of beta-gal plasmid got into the cells, then 2 parts of the luciferase plasmid should also get into the cells. Does this make sense at all?

Thank you in advance!

#2 WYF

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Posted 23 June 2010 - 06:52 AM

Hi all,

Just a quick note to say that i tried both 1:1 and 2:1 for my luciferase reporter plasmid and b-gal plasmid and both seemed to work fine, except that i am getting a very high RLU for the b-gal activity. So i am planning to reduce the amount of b-gal DNA to perhaps 10:1 of luciferase reporter and b-gal plasmids.

Encountered other problems though which i posted on another topic, with regards to the transfection being too efficient (just my speculation, may not be the case).




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