I want to setup a plate screen for a large T-DNA seed collection, where I was hoping that mutants in the pathway would grow where as the rest would show growth arrest/die similarly to a flg22 growth suppression screen. Unfortunately, I noticed that my compound photodegrades very quickly (it is mentioned in the literature and the color it shows in agar plates disapears within a few days, making then the same color as the mock plates). I then fail to see a difference between my control lines that I am using to optimize the assay. I have tried growing the seeds in plates kept in the dark but then there is no difference between the controls. Presumably this pathway requires light. I have also tried growing then in the dark for 5 days and then putting them in the light and a very small difference in growth can be seen, but it disappears within a few days, probably due to degradation of the compound. Has anyone tried to do a similar assay and, if so, what solution did you use?
Hi Daniel,
I never used this assay but just a thought. What if you try different light sources, may be a particular wavelength of light degrade the compounds quickly than others. Perhaps you could use different light filters to try.
Have you tried to protect your compaund by any black neutral pigment or membrane?
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