I am wondering if it is possible to dissolve agar by different means than boiling?
I am planning an experiment where I want to move Arabidopsis seedlings grown on ½MS + chemical inducer to plates without the chemical inducer. It is critical that I get rid of any agar sticking to the roots of the seedlings.
Would it be possible (or a good idea) to wash the roots in either water of liquid MS?
Thanks
Indeed, you could wash the roots. Arabidopsis is a weed and the roots are surprisingly tough. You could also grow the plants in liquid medium with chemical for the induction phase, so it will be easier to remove the chemical by washing. Otherwise, you can grow the plants on vertical plates, so the roots grow along the surface. How old will your seedlings be when you transfer?
In addition that Eric said, that is really good answer, you could growth the Ath in multiwells with liquid medium (depending the lenght of the assays, you can choose smaller or larger multiwells), then wash the plates and roots carfully, and put the fresh medium (if the chemical is very toxic, in my opinion should be better to chance the plate). But if you want to evaluate the effect of the chemical on root growth, the best option should be growth the seedling in vertical (or almost vertical) plate with more solid agar (because in other way, the agar is going down in the plate), then roots growth more superficial and right to down. Then you can change them to other plate (keeping sterile conditions).
Thanks for your replies. Very helpful. It is a very good point with the vertical plates.
I will grow the seedlings for 10 days and then move them to the new plate.
Regarding the liquid grown culture, I am uncertain about the specifics. Would you have the seedlings completely submerged, or would you have them suspended in a way so only the root is in liquid?
Again, thanks a lot for your answers!
The seedlings will tend to float on the liquid when they are big enough. In multiwell plates about 0.5 cm height of the liquid will be sufficient. If you need a high amount of seedlings you could also use Erlenmeyer flasks (shaking). The seedlings (roots) will lump together but it is an easy way to achieve biomass.
Fantastic!
I might just give that a try. Otherwise, I am inclined to do the vertical plates.
Thanks for the useful replies.
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