Luciferase variation among replicates

Rob Kirkpatrick kirkpat at cc.umanitoba.ca
Wed Feb 16 11:14:58 EST 2000



T Cheng wrote:

> Hi netter,
>
> I am using the luciferase assay kit to measure the luciferase activity
> of transfected primary culture of oyster cells in 96 well plate. The
> luminometer were used.
>
> However, the luciferase activity were vary among 6 replicates of the
> same treatment. The variation of luciferase activity were vary 100
> folds from 0.002 light unit (background) to 0.2 among 6 replicates.
>
> What are the possible cuases for this variation?
>
> Thanks
>
> Philip

Hi Philip,

I ran into something like this when I was optimizing my system.  It
seemed that the variability went away when I doubled my DNA that I
transfected, or really increased my DOSPER:DNA ratio (i.e. 10:1
DOSPER:DNA).  The best solution for me was to double my DNA and use a
8:1 DOSPER:DNA ratio.  Let me know how things turn out.

Rob





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