Hey,
I have a small problem.
I have cloned my target gene's UTR into pmir-Glo vector. But the problem is that frefly luciferase gene also contains miRNA target sequence. Thus when I clone my UTR back of the firefly luciferase gene, the gene contains 2 target sequence.
I will mutate the UTR sequence for the experimental procedure. Do I also need to mutate the target sequence inside the coding sequence and Do I need to use it as a non-mutated fireflyluc+UTR vector ?Or instead of that ; fireflyluc(that contains miRNA target sequence)+UTR vector can be used directly?