2 replies to this topic

[hussainmb]

Hi,

I am new to the Dual Luciferase system, so i needed some guidance. I am using Promega's dual luciferase kit.


So i did an assay transfecting cells with either

1. control TA-luc(ff)/renilla.luc or
2. TEST-luc(ff)/renilla.luc

For data analysis I divided the TEST(ff) numbers with its Renilla luc numbers.

What do i do with the control ? Do i divide control TA-luc(ff) numbers over its Renilla luc numbers, and plot it next to the test as separate bars..

OR

do i divide the TEST ratios over the control TA-luc ratios and plot as a single bar.

Any help would be appreciated.

Thanks.

[96well]

In general, I will always prefer a two bar graph. By doing so you are actually showing a control. If you normalize with the empty vector (TA-luc) it is like if you asking to the reader: 'believe me, I normalized'. If you show it, the reader will be more likely to believe the same data because his brain will normalize himself while seeing the graph.

In particular, I will not normalize twice, because the second normalization is going to delete the first one.

[Test-luc/renilla]/[TA-luc/renilla] =

[Test-luc]   [Renilla]   Test-luc
---------- x --------  = -------
[Renilla]    [TA-luc]    TA-luc

[hussainmb]

thanks a lot.. this helps..