Im am planning to do some BiFC Assays, in N. benthamiana and onion. And im wondering what kind of controls i should include.
Ideally as a positive control, I would use a gene pairs that is known to interact with each other (which i don't have) and as a negative control a gene pair that dosn't interact or has a point mutaions that prevents interaction (wich i don't have either).
Im wondering if i could just use already existing controls, that work, but have nothing in common with the genes im working with. Or if there are other ways to confirm the reliability and succhess of my experiments.
Cheers!
I have some work with BiFC and think that as a control you can use any pair of genes known to interact, independently of the nature of your genes of interest. It is a matter of interaction "yes" or interaction "no". As a negative control, you can use 1 of the positive control and 1 of your unknown set, as they are expected not to interact.
Maybe others have more ideas
Thank you for the answer! Unfortunately it came too late for me, but i used controls as you described. I still think though that the ideal negative control would be an interacting pair with point mutations, that prevent interaction, but that’s very utopic to be honest
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