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OfflineJohn in WI
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Rules of thumb for agar formulas--Are these guidelines ok? * 1
    #27086105 - 12/12/20 05:07 PM (3 years, 1 month ago)

I was digging through notes and trying to type up some things (inspired by RR's excellent cultivation notes).  Just random thoughts on things, observations, etc.

For the agar part, and after consulting FastFred's agar recipes, Frank's media journal, various books and dissertations, and the excellent Aloha Medicinal's cookbook, here are a few things that seems standard:

1)  Agar is added to water at a 1-2% rate (10-20g / liter). Sometimes a bit more for oats based agar formulas.  Lower concentration makes softer agar--often used for germination of spores.

2)  A simple sugar like malt extract, dextrose (glucose), or honey (fructose and glucose), light Karo (mostly dextrose), etc.    These are added in 0%-2% (0-20g / liter) concentrations.  Sometimes as high as 4%.

3)  Potato or other fresh root vegetables--broth made from ~200g/liter by simmering.  200g potato (with dextrose) is the standard PDA formula.  I've had good luck with 100g potato, 100g carrots (Potato-carrot agar).

4)  When using dry grain or flour, simmer 20-80g/l.  Usually 40-50g.  This was true for several cornmeal, brown rice flour, WBS, barley, and oats/oatmeal. Normally simmered for about 20 minutes and filtered.

5)  0-2g yeast.  The variety doesn't appear to matter--brewing yeast, baker's yeast, yeast extract.  Nitrogen source.

6)  The total carbohydrate level should be below 8%.  (How do you know the complex carb concentration?).  If high levels of simple sugars are used (4%), the other carbohydrate level should decrease (ie, using 100g potato instead of the regular 200g for PDA).

7)  The carbohydrates concentrations can alter mycellium morphology.  Using lower concentrations can favor vigorous/rhyzomorphic growth as the fungi stretches to find nutrients.


A few questions--why wouldn't you just the same carbs that are in the final substrate?  Ie, PDA (which works great for growing mycellium).  The final substrate would never be potatoes.  Is it a good idea to go through PDA, vs. oatmeal, corn, or WBS based agar.

Anyone else want to contribute rule's of thumb for formulating agar?  Thanks for any insights or corrections


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OfflineBrian Jones
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Re: Rules of thumb for agar formulas--Are these guidelines ok? [Re: John in WI]
    #27086298 - 12/12/20 07:10 PM (3 years, 1 month ago)

I get confused when the measurement is too scientific.
In each 500ml media bottle I use 8 grams of agar agar, 8 grams of light malt extract and 400 ml of water.


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InvisiblePastywhyteMDiscord
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Re: Rules of thumb for agar formulas--Are these guidelines ok? [Re: Brian Jones]
    #27086308 - 12/12/20 07:17 PM (3 years, 1 month ago)

I love PDA. Works great for getting invitro pins.


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OfflineJohn in WI
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Re: Rules of thumb for agar formulas--Are these guidelines ok? [Re: Pastywhyte]
    #27086687 - 12/13/20 04:20 AM (3 years, 1 month ago)

That's good info.  I was cleaning up some old dishes, and found a couple that were full of knots, small pins, and in a couple of cases fully mature and spore dropping fruits.

It seems to me, if you are getting knots and pins on agar that you must certainly have a fruiting strain?  I have a photo someplace--one of the dishes had a knot every 1/8".  Just covered with them.  I transfered a bunch of them to new plates and want to grow them out.


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InvisibleMadHatter333
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Re: Rules of thumb for agar formulas--Are these guidelines ok? [Re: John in WI]
    #27086705 - 12/13/20 04:52 AM (3 years, 1 month ago)

I always got great results using 8g agar 8g brf to a hair over 400ml of water. I always got great rhizo growth that would practically jump off the plate. I tried malt and that sucked for me and the growth was weak. Definitely like brf though!


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OfflineJohn in WI
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Re: Rules of thumb for agar formulas--Are these guidelines ok? [Re: MadHatter333]
    #27087108 - 12/13/20 10:48 AM (3 years, 1 month ago)

This is something I'm trying to learn about.  Like using BRF as an agar nutrient, even though you wouldn't (probably) use brown rice grain as an actual grain spawn. 

Also, it seems that oat or oatmeal based agar is excellent for any of the pan species.  It's curious why the carbohydrate profile has such an influence on the growth speed, and the resulting morphology. 

For example, I'm diabetic.  The rules are fairly simple--simple sugars (glucose, fructose, sucrose...) spike the shit out of my sugar.  My body can't really tell the difference.  Complex carbs, whether they come from brown rice, whole wheat, etc?  They work at a much slower rate (as enzymes have to break it down first).

I just bring it up because in my experience, colonization rate on whole grain is much more a function of proper hydration and number of inoculation points than in the actual composition of the spawn.  Popcorn goes fast, whole oats, WBS...  Cubes really don't seem to care that much (again, assuming hydration is right).

It's interesting stuff.


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