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InvisibleUna
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Registered: 03/01/01
Posts: 970
Some proven agar nutrient media recipes
    #3033489 - 08/21/04 07:12 PM (12 years, 6 months ago)

Here are some media recipes that support healthy mycelial growth of the following species:
Psilocybe cubensis, Ps mexicana, Ps tampanensis, Ps. azurescens, Ps. cyanescens, Panaeolus cyanescens, Pan. tropicalis, Pan subbalteatus (and many more...)

These are all media i have been using myself for the past couple of years.

Malt Extract Agar

(for 1 liter)
20 g extra light malt extract
2 g yeast
15-20 g agar

Put dry malt extract, yeast and agar in a bottle, add water, swirl,sterilise.

Malt Yeast Peptone Agar

(for 1 liter)
20 g extra light malt extract
1 g yeast extract
1 g peptone
15-20 g agar

Put dry malt extract, yeast extract, peptone and agar in a bottle, add water, swirl, sterilise.

This medium is a bit richer than standard MEA and some of the ingredients are somewhat harder to buy for indivuals.

Potato Dextrose Agar

Boil 200 grams of sliced potatoes for 30-60 minutes, strain out the potatoes save the broth. Dissolve 20 grams of dextrose in the broth. Put agar in a flask, add the broth and fill out to 1 liter with water, sterilise.

A very good allround medium which i hardly ever use. It just is too much work to prepare.....

Oatmeal Agar

Combine 75 grams of oatmeal flakes with 1/2 liter of boiling water. Stir for 5-10 minutes then filter out the larger particles by pouring it through some mesh, save the broth. Put 20-25 grams of agar in bottle, add the oatmeal broth, fill out to 1 liter, swirl, sterilise.

This is the best medium for Panaeolus cyanescens i've ever encountered. Beware, this medium will be less firm than the other recipes so extra agar has to be added to compensate.

Smaller quantities of any media can be prepared simply by using less ingredients in the right proportions.

When making your own recipes keep in mind that more is not always better. When media are too rich in nutrients mycelium will not grow or grow very poorly secreting yellowish metabolites.

In my experience tap water works just as well as distilled water but then again, we have pretty good chlorine free tap water here. If in doubt, better to use bottled water

Sterilisation Tips

Agar normally is sterilised in bottles or erlenmeyer flasks. Since agar foams during autoclaving the flasks are never filled for more than 2/3. Bottles with water can be totally filled. The caps of bottles or flasks should never be screwed tight but always left a bit open to prevent im- or explosions.

It's very important that the cooker cools down slowly. When the temperature drops too fast the agar may boil out of it's container which leads to mess.
When agar media are sterilised too long they turn dark in color and don't gel properly. Mycelium does not grow properly on such media and often shows sectoring.

Sterilising culture tube slants

The nutrient medium is prepared by bringing the appropiate amount of water to a near boil in a pot on the stovetop. The ingredients are added and the mixture is stirred to let them dissolve. Agar should first be mixed with a little bit of cold water before adding or it will form lumps. Careful, adding agar to boiling water will cause the medium to foam and boil over. Boiling agar can cause serious burns!
Let the agar boil for a few minutes, then turn off the heat.Tubes are filled with 5-6 ml of medium and loosely capped. A small funnel and a big syringe are very useful here. The tubes are sterilized for 15-25 minutes at 121?C. When they come out they are left to cool on a sloped surface to increase to surface area of the agar. When the agar has solidified the slants are ready to use.

Sterilisation time
There is much discussion about how long agar medium should be sterilised. One thing left out of this whole discussion is the container volume.
Proper sterilisation time depends on the amount of medium in one container. A testtube with just a few milliliters is sterilised for 15-25 minutes. A bottle with 500 ml is sterilised for 35 minutes. Insufficient sterilisation will lead to problems with bacterial contamination. Over sterilisation will lead to darkening of the medium commonly referred to as 'caramelisation' and a drop in pH which leads causes the agar to break down so it won't gel anymore.


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InvisibleCorporal Kielbasa
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Re: Some proven agar nutrient media recipes [Re: Una]
    #3036140 - 08/22/04 01:54 PM (12 years, 6 months ago)

Thanks for the write up. When sterilising medium I like to stuff a big wad of papper towel in the mouth of the flask.

If using grolsh beer bottles never close the cap. This could lead to explosions or a guiser like propultions of media.


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OfflinePsilygirl
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Re: Some proven agar nutrient media recipes [Re: Corporal Kielbasa]
    #3036155 - 08/22/04 01:57 PM (12 years, 6 months ago)

just use foil (make sure its autoclavable!)


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InvisibleCorporal Kielbasa
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Re: Some proven agar nutrient media recipes [Re: Psilygirl]
    #3036245 - 08/22/04 02:36 PM (12 years, 6 months ago)

I like how the papper towel makes a nice obsorbent plug. But foil works well also. I try to make a lil more then a litre at a time so I push the edge of safety when using my flask.


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Invisiblefastfred
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Re: Some proven agar nutrient media recipes [Re: Corporal Kielbasa]
    #3036550 - 08/22/04 04:45 PM (12 years, 6 months ago)

Foil is the standard method. You shouldn't push the limits on flask volume when making media, unless you really know what you're doing. Media is a lot different than plain water and will boil over much easier.

The rule of thumb is to fill your flask half full. And certainly no more than 2/3 full.


-FF


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Offlinedjred
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Registered: 08/27/04
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Re: Some proven agar nutrient media recipes [Re: fastfred]
    #3145344 - 09/16/04 09:48 PM (12 years, 5 months ago)

hmm..i've been stuck in the basic growing rooms lately, and to come in here as researching ideas.your telling me i can use some plain old house hold items to makea liquid medium for the myc to grow on.

well i take it then they still need to be cased or given some kinda of substrate to grow on? well guys i enjoy reading all these neat shit you guys come up with ..great site..i find somehting new everyday..love it...


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OfflineErik006
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Registered: 07/13/03
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Re: Some proven agar nutrient media recipes [Re: Una]
    #3149663 - 09/17/04 09:03 PM (12 years, 5 months ago)

>When the temperature drops too fast the agar may boil out of it's container which leads to mess.

Agar does create a mess :wink:

In my begin phases of cultivation I dropped a 2/3's of a liter of agar, and it was hard to get off my washing machine and carpet.

But thats besides that point, For sterilization I now use a quart jar filled about half way, this usually gives me enough agar to fill three petri dishes. (note, I own large size petris), I sterilize the agar and dishes at the same time for about 25-30 mins. And fill in front of my flow hood.

Great write up Una, it is  very much appreciated!

Erik006


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OfflineAnnoA
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Re: Some proven agar nutrient media recipes [Re: Erik006]
    #3151034 - 09/18/04 08:22 AM (12 years, 5 months ago)

>a quart jar filled about half way, this usually gives me enough agar to fill three petri dishes.

500 ml for 3 petri dishes?!
How big are those petri dishes??


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InvisibleCorporal Kielbasa
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Re: Some proven agar nutrient media recipes [Re: Anno]
    #3155292 - 09/20/04 12:52 PM (12 years, 5 months ago)

I hope he ment three sleeves. 

Has any one every had the privelage of cleaning agar of the ceiling and walls?  Its not fun at all, that why I had my girl friend do it. :smile:


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OfflineAnnoA
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Re: Some proven agar nutrient media recipes [Re: Corporal Kielbasa]
    #3155325 - 09/20/04 01:02 PM (12 years, 5 months ago)

>I hope he meant three sleeves.

500 ml fills 1 sleeve (20 dishes) here.....


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InvisibleCorporal Kielbasa
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Re: Some proven agar nutrient media recipes [Re: Anno]
    #3155348 - 09/20/04 01:09 PM (12 years, 5 months ago)

How many mills in a litre? 1000 right? I think I have done 4 sleeves to the litre. But they poured mighty thin


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OfflineYSr
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Re: Some proven agar nutrient media recipes [Re: Corporal Kielbasa]
    #3209668 - 10/03/04 03:15 PM (12 years, 4 months ago)

Hmmm... liquid mediums (dextrose or ME) call for about 4% sugar or malt, but
why only 2% ME in ME agar?

YSr


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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