Ok, I have been using a 2-3% malt concentration in my agar for a long time, and have had no trouble, finally settling on an additional 0.5% concentration of Dextrose as well. Now since I have decided to use rye or straw in my agar as well (typically 0.5% for each supplement), It has me wondering about concentrations of such ingredients when combined. Typical recipe is: 200ml water, 4 grams malt, 1 gram dextrose, 1 gram Yeast, 1 gram powdered substrate or 1ml of a true substrate extract.
I am curious to know when you would start to run into diminished returns adding additional supplements, I.E. - at what levels would start to hinder growth. What happens if I make a 10% malt concentration for example? Is there an upper limit (or an ideal limit) to where I should balance my total sugar and carbohydrate concentration?
In TMC Stamets mentions an agar mix utilizing boiled rye supplemented with malt, but does not go into further detail. Basically I would look at it like a recipe I used in reverse, 4 grams malt to 1-2ml rye extract. Anyone have a suggestion on where I would start with this? I have no clue how much rye to water ratio to use when boiling, how long, or how much malt to add. Stamets likes to leave us hanging I guess or more importantly, figure it out on our own (which I don't want to do, I have enough experiments going as it is).
Next question: One thing I don't think I have ever seen covered is how "solid" should the agar be? I personally have had different brands and some premixes come out hard like wax almost, while others are just like jello. I also notice Stamets mentions when making slants he "doubles up on the formula so the agar gels sufficiently" (GGMM, I assume he only means agar). I use 5 grams agar for my dish mix, and this usually leaves me with an agar much harder than jello, but still soft and squishy. I add an additional gram when I make culture tubes, usually always about 6-7. It becomes quite a bit firmer than jello, almost rubber like. What is an ideal "hardness" level, or does it really matter that much? Will the mycelium still colonize it just as well if I were to increase up to 8 grams agar?
Last question: I am at the stage where I would like to alternate my media recipe for each transfer, helping with mycelial vigor and reducing media selectivity. I can definitely notice the leap-off time is reduced by at least a day using rye extract in my agar when transfer a wedge to a jar. I have a curiosity that since the mycelium was currently digesting nutrients in the grain agar, it had no problem grabbing on to a rye kernel. But what about later transfers, say I use OMYA with no grain. This "new" mycelium would not currently be in the process of digesting rye nutrients, but would it still retain a "memory" for the substrate, thus recovering and taking to the grain in a shorter time as I have observed? I ask that because I wonder if I am wasting my time powdering straw for agar that gets transferred to grain...
Ok wait, one last quickie: Is peptone worth the extra time to weigh it out?  I have 25g on the way to try it out, which is more than enough to last me for awhile. Does it help with the vigor in a noticeable way?
Sorry for the long wind, its my nature I guess...
-------------------- To give is to live...
|