Hello everyone!
I have a question regarding deselecting plasmids from yeast transformants. For episomal plasmids, culturing transformants in non-selective media several rounds can do the trick. However, I have not yet found a method that can make yeast lose their CEN/ARS plasmid? Does anyone know a method that can achieve that?
Thanks!
Non-selective growth also works for centromeric plasmid you may simply have to pick more colonies. Alternative to that, For plasmids carrying the URA3 marker you can use 5-foa which is toxic if the cells carries a functional URA3 marker. For plasmids carrying TRP-1 marker you can use 5-fluoroanthranilic. For plasmids carrying Met17 you can use color based screening with lead sulfide this lead a dark coloration for cells that lost the plasmid. For plasmids carrying either an antibiotic marker or other auxotrophic marker you may wish to add another marker for counterselection like amdSYM (amdSYM, marker can be counterselected with fluoroacetamide and can also be selected with growing cells with acetamide as sole nitrogen source).
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