I need to make plates of fission yeast YE5S medium with 1.2 M sorbitol. An old protocol in the lab says to make 0.5 L of 2.4 M sorbitol as well as 0.5 L YE5S + enough agar for 1 L, autoclave separately, then mix and pour plates. I am unsure of several things.
1) Shouldn't the amounts of YE5S components also be enough for 1 L, since that is the total volume I will eventually end up with? As is, the recipe will produce half-strength YE5S with 1.2 M sorbitol.
2) Why autoclave the sorbitol separately? Sorbitol (from what I can find) doesn't appear to undergo the Maillard reaction, which I believe is the primary reason for autoclaving sugars separately when making media. Additionally, if that is the concern, shouldn't the dextrose also be autoclaved with the sorbitol, instead of the rest of the YE5S components?
Hi Sam,
you definitely need to use double-strength YE5S to mix it in a 1 by 1 ratio with 2.4 M sorbitol. However, I cannot come up with an answer to the sorbitol/glucose question. It is not too much trouble to autoclave sorbitol and medium separately so I would do it.
Best wishes
Jürgen
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