I have made an ethanol inducible miRNA construct for knock-down expression of my protein of interest to detect phenotypes in seedling growth. Currently I'm growing the seedilngs vertically on hard agar and treating with 2% ethanol by pipetting onto the agar and roots twice a day. I'm seing about 30-50% reduction at the protein level, but would like to increase that to around 100%. However, I feel if i pipet any more liquid onto the plates, the seedlings will become water-logged.
Is anyone else doing ethanol induction in seedlngs? If so I would love some suggestions on how you are inducing you protein or RNA of interest.
University of WI - Madison