Hello all,
I am wondering if there is any cheap method for plant cell wall disruption and getting DNA, RNA and proteins into solution. Obviously, there are many standardized ways:
Liquid nitrogen grinding
Beads milling
Enzymatic digestion of cell wall, etc.
But I am interesting in some alternative way, ideally feasible with common laboratory reagents. Does anyone know such a low-cost method how to destroy plant cell wall ? :-)
Thanks for your opinions
Martin
@ Sreetama Bhadra,
subsequent RNA and protein isolation, probably using Trizol
Hi, if you are interested in cheapness, you have also to consider waste management (which can also be considered inn some cases expensive as well). Of course it also depends on what you are interested in (nucleic acids, protein, everything?). You can consider sonication, blending, that are preparatory for chemical-mediated stabilization of the matrix, like phenol:choroform
Liquid nitrogen in combination with tissue crusher/cell homogeniser. Since your downstream targets encompassing RNA, DNA and proteins, which are highly susceptible to temperature, proteolytic and phosphorylation. Achievable at laboratory scale and reproducible with consistency. All best luck!
Your aim is to isolate DNA, RNA and protein. The 'Enzymatic digestion of cell wall' method is not a good choose. Enzyme reaction usually needs a higher temperature, such as room temp (25C). Besides, some protoplasts (no cell walls) break after cell-wall enzyme digestion. Broken protoplasts release all kinds of nucleases which readily destroy DNA and RNA, especially at room temperature. Enzymatic digestion of cell wall is useful for 'protoplast isolation'.
Liquid nitrogen and ball milling methods are performed under cold temperature, so cause less problem.
@ Stefano D'Alessandro,
"Regarding proteins you can simply add plant material to the loading page and boil it."
Won't this approach denature the proteins by the heat? We used to isolate proteins at 4C environment, and added anti-protease agents in the extraction solution.
this link is useful
https://www.thermofisher.com/...methods/traditional-methods-cell-lys...
regards
Follow this link maybe useful
https://bitesizebio.com/13536/bringing-down-the-walls-part-ii-8-methods-to-break-down-cell-walls/
regards
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