- I'm doing a stable expression of dual luciferase assay system with one vector. But I'm getting low firefly fluorescence value, around 300 to 800, and my renilla fluorescence value is around 10000 to 20000. I’m wondering whether the firefly fluorescence value is lower than the detection limit of the Promega Glomax 20/20 machine and whether the results are reliable.
- Thank you in advance.
Yuning Hou
You got the Renilla luciferase activity means the reaction system is working, and the 300-800 RUL value should fall into the detection range of the reader. Usually, the Rirefly luciferase activity is expected to be comparable to or even higher than Renilla luciferase. As you are using one vector to express two different reporters, I assume that different promoters regulate the luciferases.
So, the first question is, do these two promoters possess the same efficiency in driving the enzymes' expression?
The second question is, do you have a negative or positive control? If so, what are the RULs for the controls? If not, I would suggest including control groups.
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Physical Phenomena
- Luminescence
- Fluorescence
More Yawen Xiao's questions See All
Similar questions and discussions