Hello,
I'm growing different accessions of Arabidopsis thaliana and so far we have been harvesting them by pouring the media over a simple sieve, then putting them on paper towels for a minute to try to remove excess media. However, too much still remains and the samples become total rocks in the freezer and it makes metabolite (MeOH) extraction really difficult.
Is it okay to use soft vacuum filtration for a few seconds to harvest the seedlings? Which type of membranes can I use with 0.5 MS (pH 5.7) media? Our department generally stocks PES.
Or maybe I should do the extractions right away?
Thanks,
Casey
Hello, Casey!
In no case you could not do the MeOH extraction while there is some media in your seedlings. The sugars in your media will introduce changes in content of them in plant extract.
I really don't know how to help you in situation you are in, but in the future you can use more solid media and grow seedlings in vertical position of Petri dishes, so roots will not penetrate it very much.
Good luck!
Hi Casey
I am not familiar with the method, but here is an idea, which just occured to me:
We are using a kind of mesh for extraction of Protoplasts.
This comes also as a so called strainer (https://us.vwr.com/stibo/bigweb/std.lang.all/74/48/13587448.jpg), where you could pour the seedlings on and then maybe rinse with some ddwater, before drying them.
And then maybe you could try vaccuum on this, as there should not be any sugar on them any more...
Hi Casey,
I think you can try and find out. I think general Whatman membrane (filter papers) is good enough. You can rinse the tissues with clean water while you gentle vacuum them, both the water and residual medium will be removed through the gentle vacuum.
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