Hello researchers,
For now, I have developed PCR-confirmed and antibiotic-selected stable transgenic rice plants. I was doing qPCR expression analysis for my gene of interest, but I am getting no expression in my GOI. The selection marker is showing expression in the main experiment. During the trial, GOI expression and the antibiotic selection gene were present. We thought primers might have degraded, and we brought new primers even though it's not showing expression. What might be the reason? Have you come across any such problem? Please help me with this.
Thank you
Supriya
Have you done appropriate controls to determine whether the problem is that your samples themselves or the PCR? In other words have you done PCR controls with some other sample or a very tiny amount of the plasmid to show that is still working. Even better would be to use add the plasmid to your plant DNA sample and confirm there is nothing inhibitory.
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