What are the purposes of the various metals in the solutions used in protoplast isolations? In the digestion solution, 0.2 mM CaCl2 is used (which seems hardly anything, but may be important for the cellulase and pectinase enzymes). In the stop solution (W5), 125mM CaCl2 is used (which I assume is to help stabilize the membrane) and 5mM KCl is included (what is the purpose of this?). Then, in the MMg solution (aptly named because it has 15mM MgCl2 in it), with no other metals. After that, the cells are incubated in WI, which has 20mM KCl in it. 1. Why are these particular metals used in these specific solutions and 2) why do the species change and 3) why do the concentrations change? I want a better functional understanding of this process, and I’ve looked in the literature, but I assume this is so far back, and was “common knowledge” that I was not successful in finding an explanation. Any input is appreciated.
I think that adding the mineral elements or positive ions in a salt form such as CaCl2 is to reduce the value of zeta-potantial electrostatic on the surface of the protoplast, which is usually charged with a negative charge, which leads to repulsion of cells and prevents their convergence during the process of electrical or chemical fusion ... Therefore, positive ions such as Ca ++ On the negative charge equation on the surfaces of the protoplast and convergence allow it to reduce the value of zeta-potontial
Hi Taha. That's a very good explanation for the calcium. Do you know anything about the metals in the other solutions besides the transfection reagent? For example, why does MMg have magnesium in it, while W5 and WI have KCl in it? Why the switch?
Actually I have some information about zeta-potential, but currently I do not have knowledge of the rest of the question and thank you for your response
Hi Dr.Monique, can you tell me about Ideal answer for your question؟؟
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