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InvisibleATWAR
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Re: FastFred's Media Cookbook [Re: ]
    #3025865 - 08/19/04 09:12 PM (19 years, 10 months ago)

Some people just don't get the point...  :rolleyes:



Now that the thread has again become productive, for newbies I would personally recommend you go with an agar premix available from the Shroomery sponsors such as  The Spore Works. I started out making my own MEA with ingredients purchased from a health food store with no problems. I still have the two pound bag of malt sugar I purchased to this day (although it is considerably smaller)...

I tried a few different premixes also, but I will stick with making my own. It is cheaper and you can alter the concentrations. Premixes work well and are forgiving for the newbie. You only have two things to measure - Agar mix and water. It really doesn't get much easier than that...


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Invisiblefastfred
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Re: FastFred's Media Cookbook [Re: ATWAR]
    #3025935 - 08/19/04 09:27 PM (19 years, 10 months ago)

I recommend the PDA, or MEA recipes in the cookbook. They are both easy to make and have easily obtainable ingredients. PDA and MEA both perform about the same. The PDA might be slightly more resistant to contamination.

As always, support the shroomery sponsors. But you should be able to get everything you need besides the petri dishes or jars from your local healthfood store. Dextrose is corn sugar. They usually also have food-grade agar. One warning though, food-grade agar usually has a lower gell strength than lab-grade agar, so use slightly more agar. 20g instead of 15g or 25g instead of 20g. It won't affect the quality of the agar. If it gels too strongly simply use a little less agar next time.


-FF

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InvisibleATWAR
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Re: FastFred's Media Cookbook [Re: fastfred]
    #3026039 - 08/19/04 09:54 PM (19 years, 10 months ago)

As for foodgrade agar, if you have a choice try to get this brand:


It is the best I have ever used. Sometimes you can get agar from bulk bins at healthfood stores but I don't recommend you use that stuff. This brand works very well for me, just as well as media grade agar. It will gel sufficiently at 1.5% (15 grams per liter) although I prefer mine a bit harder. At 1.5% it will come out a bit harder than Jello (2% is generally standard, but I sometimes use up to 3-4% for slants)...

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InvisibleSpeeker

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Re: FastFred's Media Cookbook [Re: fastfred]
    #3026972 - 08/20/04 01:55 AM (19 years, 10 months ago)

on +40 minutes sterilization times:

Maybe Stamets has become a bit over-cautious because of the problems he had..

He discuss about it here:
http://www.ibiblio.org/pub/academic/agriculture/sustainable_agriculture/mycology/bionet.mycology/

(the big file, "Bionet.mycology", ~5MB)

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Offlineliveby
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Re: FastFred's Media Cookbook [Re: fastfred]
    #3027002 - 08/20/04 02:10 AM (19 years, 10 months ago)

I did pasta and rice!

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InvisibleUna
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Re: FastFred's Media Cookbook [Re: fastfred]
    #3031793 - 08/21/04 07:27 AM (19 years, 10 months ago)

@fastfred

What's the point of posting so many recipes when most of them will not support proper growth of mushroom mycelium. You back everything up with literature references instead of own experience. Have you ever yourself made agar medium?

Quote:

I hope you guys will grow up and start putting some content in your posts rather than just bitching about the people who are actually adding content to this board.



Yes, pretty useless content if you ask me. Again, posting a list with so many recipes and not telling that some of them are useless i consider misinformation.

Quote:

I'm just telling you guys the standard wisdom here... And out come people calling me a fool and asking for citations! Why don't you guys come up with some citations, you are the ones telling me information that is contrary to every bit of literature that I've read.




I don't need citations i speak from experience which i think is a lot more valuable than the result of a 10 minute Google search, any fool can do that...Now how about you tell us which media worked best for you.

At this point i am assuming you have never grown a mushroom and are trying to impress everyone with your book-knowledge. I guess some people just don't care how they get attention as long as they get attention.....

Now to add some USEFUL CONTENT

Best media for mushroom mycelia: MEA, MYPA, PDA and OA

The proper sterilisation time depends on the volume of the container the medium is sterilised in. Test tubes will be fine with a 15 minute sterilisation but don't expect a bottle with half a liter to be sterilised in the same amount of time. A bottle like that will need 35 minutes to be completely sterilised.

In the whole discussion about the proper sterilisation time, the container size/volume is totally forgotten.


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Invisiblefastfred
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Re: FastFred's Media Cookbook [Re: Una]
    #3036789 - 08/22/04 04:37 PM (19 years, 10 months ago)

Quote:

Una said:
What's the point of posting so many recipes when most of them will not support proper growth of mushroom mycelium. You back everything up with literature references instead of own experience.




AFAIK all of the media types listed will support proper growth of mycelium. Are you even reading this thread? I already posted that I would revise the guide if anyone has experience with any of these medias not working. That was in response to your same assertion earlier that some of them would not work. So if you know of any that won't work, post it... Otherwise shut your mouth.

The reason I cite references is to give proper credit and assist anyone who wants to do further research on the subject. It's also a lot nicer to be able to cite a textbook rather than having to say "an unknown author at the Shroomery". This makes it a lot easier to write up your experiments in a normal fashion.


Quote:

Have you ever yourself made agar medium?




Hundreds of liters.


Quote:

Again, posting a list with so many recipes and not telling that some of them are useless i consider misinformation.




Well, since none of them are useless, your point makes little sense. If you've ever looked at a list of media types you should realize that there are hundreds of standardized types of media. My idea was to reduce this by eliminating ones that won't work or are complicated to make. I think I've done pretty well, and I plan to expand the cookbook significantly with non-standard recipes in the future.


Quote:

I don't need citations i speak from experience which i think is a lot more valuable than the result of a 10 minute Google search, any fool can do that...Now how about you tell us which media worked best for you.




Again, are you even reading the thread? PDY, MEA, and PDA are the best types to get started with. I already stated this just a few posts back.


Quote:

At this point i am assuming you have never grown a mushroom and are trying to impress everyone with your book-knowledge. I guess some people just don't care how they get attention as long as they get attention.....




You assume wrong. I've grown several types of edible and medicinal mushrooms. I'm not trying to impress anyone. I'm just trying to clean up the sloppy state of the science here at the shroomery.


Quote:

Now to add some USEFUL CONTENT
Best media for mushroom mycelia: MEA, MYPA, PDA and OA




MYPA? Do you have a citation, or is it just something you made up yourself?


Quote:

The proper sterilisation time depends on the volume of the container the medium is sterilised in.




The quoted time of 15-20 min is for 1L of media. I think that is pretty clear in the cookbook.


Quote:

Test tubes will be fine with a 15 minute sterilisation but don't expect a bottle with half a liter to be sterilised in the same amount of time. A bottle like that will need 35 minutes to be completely sterilised.




Liquid media does not require the extended sterilization times required for substrate. 15-20 minutes is fine for 1L of media.


Quote:

In the whole discussion about the proper sterilisation time, the container size/volume is totally forgotten.




No, it is not forgotten. The specified volume is 1 liter, the specified sterilization time is 15 min. Did you even read the guide?

Please post again when you've actually read the thread and have somthing to contribute.


-FF

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Offlineragadinks
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Re: FastFred's Media Cookbook [Re: fastfred]
    #3036846 - 08/22/04 04:54 PM (19 years, 10 months ago)

> In the whole discussion about the proper sterilization time, the container size/volume is totally forgotten.

I think this whole point is a question of definition:
When it says: stereilize 15 minutes at a temp of 121 degrees Celsius than it means that the *whole* substrate has to have a temperature of 121 degrees for at least 15 minutes.
When you say that a larger container needs longer sterilization times you probably consider the time to heat the whole medium up to 121 degrees Celsius to be within the sterilization time ?
I do not think that a larger volume of substrate needs longer sterilization time.
It only takes much longer till all the medium is heated to the proper sterilization time (also inside of it - not only on the outside ).
Not sure if I have expressed myself properly - hope my point is clear ?

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Invisiblefastfred
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Re: FastFred's Media Cookbook [Re: ragadinks]
    #3036880 - 08/22/04 05:04 PM (19 years, 10 months ago)

> When it says: stereilize 15 minutes at a temp of 121 degrees Celsius

It means exactly what is says. You load 1 L of medium into your autoclave, when the temp reaches 121C your start the timer and cook for 15 min. There is no measurement of media temp. Liquid media heats quickly and evenly, unlike solid substrate.

It may take 30-45 min for the entire cycle, including heat up and cool down times, but you only count the time that it is at full temperature.


-FF

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InvisibleATWAR
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Re: FastFred's Media Cookbook [Re: fastfred]
    #3047230 - 08/25/04 01:26 AM (19 years, 9 months ago)

Quote:

fastfred said:
I'm just trying to clean up the sloppy state of the science here at the shroomery.




Whoa there... In case you didn't notice we already have media recipes listed in both the FAQ, and the cultivation guide. If you think the Shroomery is sloppy, feel free to leave. I don't think you will be missed...

Quote:

MYPA? Do you have a citation, or is it just something you made up yourself?




I will speak for UNA on this one. That would be malt yeast peptone agar. The most common recipe can be found in GGMM by Paul Stamets.



I am curious to know if you have grown many edible and medicinal mushrooms why you think that some of the recipes that were not originally developed for mycelium growth (but rather bacterial growth) are useful to us? Many of these recipes are questionable IMO, particularly the following:

Plate Count Agar (the name of this one implies it is not for mycelia)
Nutrient Agar (what type of mushrooms favor beef?)
YE - Yeast Extract Agar
GA - Gelatin Agar

And then you cite your resources for the above recipes: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998. Your source for these recipes also implies they were not developed for use with mushroom mycelium but rather bacteria, yeasts, and molds.

But before I say these are useless, I will put them to the test so I can speak from experience. While I have no doubt mycelium will grow on these, it is my guess they will not perform nearly as well as the more common recipes. Then again I could be wrong, and I plan to find out...

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InvisibleUna
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Re: FastFred's Media Cookbook [Re: fastfred]
    #3052019 - 08/26/04 03:39 AM (19 years, 9 months ago)

Thanks ATWAR, couldn't have said it any better :laugh:

@fasfred

Since you like citations and references so much i took the trouble to find you this. Yes, i know, it refers to plant tissue culture media, but that's totally irrelavant here. It's a table showing the required autoclaving time for different volumes of media.

http://www.sigmaaldrich.com/Area_of_Inte...rilization.html



Still you haven't told us which of these exotic recipes (not MEA, PDA) work best for you?


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Edited by Una (08/26/04 07:53 AM)

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Offlineragadinks
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Re: FastFred's Media Cookbook [Re: Una]
    #3052036 - 08/26/04 03:53 AM (19 years, 9 months ago)

Thanks UNA for that citation - it expresses very much the point I was raising:

Quote:

Minimum autoclaving time includes the time required for the liquid volume to reach the sterilization temperature (121?C) at 15 minutes.



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Invisiblefastfred
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Re: FastFred's Media Cookbook [Re: ragadinks]
    #3054610 - 08/26/04 05:53 PM (19 years, 9 months ago)

That's not really an issue. As I said earlier, liquid media heats quickly and evenly.

If you're using a pressure cooker then your glass media container is probably sitting actually in the water, so your chamber temp and vessel temp will be nearly identical.

Any lag time in warming up the media is more than compensated for by the cooldown lag time.

15-20 min at 121C is fine for media volumes of one liter or less. Larger volumes should be separated into 1L or less amounts. Using half the volume of a standard quart jar, your media volume per vessel will be around 1/2 L(500 ml).


-FF

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InvisibleUna
controlleddemolition

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Re: FastFred's Media Cookbook [Re: fastfred]
    #3062808 - 08/28/04 05:21 PM (19 years, 9 months ago)

That's not true. Larger volumes DO need longer autoclaving. This is getting silly, why are you so persistent?

Stamets is right, I found this out myself during my studies, before i ever poured a dish with MEA, PDA or any other medium intended for mycelium.
I lost hundreds of tissue culture plants due to contaminated medium. The 20 minute sterilisation cycle that worked fine for the medium filled tubes was too short for 1 liter bottles with 0.5 liter medium (i was moving the plants to plastic boxes).

I have autoclaved a lot of medium, for mycelium as well as for tissue culture plants. In the few cases that totally contaminated medium was encountered it could always be traced back to a wrongly set timer clock (setting the correct time for tubes instead of bottles).

Here's some interesting reading material about the subject.

http://www.rbgsyd.gov.au/conservation_re...tion_techniques

An effort should be made to avoid sterilising large and small volumes of media in one load as time must be allowed for large volumes to reach the required holding temperature, and this will result in small volumes receiving too much heat. Here is a rough guide to the extra time that must be added to reach holding temperature:

Volume of liquid
Extra time
Total time at 121?C

100 mL bottle
10 mins
25 mins

100 mL conical flask
2 mins
17 mins

250 mL bottle
12 mins
27 mins

250 mL conical flask
4 mins
19 mins

500 mL bottle
18 mins
33 mins

500 mL conical flask
8 mins
23 mins

1000 mL bottle
22 mins
37 mins

1000 mL conical flask
12 mins
27 mins

2000 ml bottle
27 mins
42 mins

2000 mL conical flask
20 mins
35 mins

http://www.800ezmicro.com/microBiology.asp?mb=88

Always remember that the temperature inside the AUTOCLAVE vessel and the MEDIA vessel will be different until equilibrium is reached. The actual sterilization time is only a fraction of the total cycle time. The heating and cooling phases also take much longer time than the actual sterilization time. When the autoclave vessel reaches sterilization temperature, the liquids still have a lot more energy to absorb before they reach sterilization temperature. As a result the overall time required to attain equilibrium between chamber and liquids is much longer.

http://www.esd.uga.edu/bio/Autoclave_Ops.html

Average liquid sterilization times (add an additional 10 to 20 minutes for crowded items):
<500 ml<>,
30 minutes
500 ml - 1 L,
40 minutes
2 L - 4 L,
55 minutes
4 L,
1 hour

http://www.phytotechlab.com/TABSTYLE/webdocs/Sterilizing%20Nutrient%20Media.pdf

For small volumes of liquids (100 ml or less), the time required for autoclaving is 15-20 min, but for larger quantities (2-4 liter), 30-40 min is required.


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InvisibleATWAR
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Re: FastFred's Media Cookbook [Re: Una]
    #3063314 - 08/28/04 08:14 PM (19 years, 9 months ago)

It should be common sense that the volume of media dictates its sterilization time. There are other variables to take into account as well such as size and shape of the vessel, and material it is constructed from. When a PC first reaches full pressure, the media inside the flask will not be at 121C. There is a lag time that must be compensated for. My sig should explain the problem here...


And for the media debate, I have just finished inoculation of the following with four different species:
Nutrient Agar
Yeast Extract Agar
Gelatin Agar
MnK Agar (milk and cookies)
MYPA (control)

I figured since I was trying obscure media recipes, I would include one of my own. So naturally when I was eating milk and cookies, the idea just hit me. I made milk and cookies agar... We shall see in a week or so how these media recipes compare to a standard media recipe that has been proven to work very well with mushroom cultures...

Unil then,
ATWAR


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To give is to live...


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Offlinekilgore_trout
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Re: FastFred's Media Cookbook [Re: ATWAR]
    #3063424 - 08/28/04 08:56 PM (19 years, 9 months ago)

oreos, chips ahoy, homeade?

are you leaving out that you were HIGH and eating cookies?

a word of advice for next time, "a cookie is a cookie, but newtons are fruit and cake."

let us know how it turns out.


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InvisibleATWAR
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Re: FastFred's Media Cookbook [Re: kilgore_trout]
    #3063863 - 08/28/04 11:22 PM (19 years, 9 months ago)

Quote:

kilgore_trout said:
are you leaving out that you were HIGH and eating cookies?




No. I was just hungry. I was thinking about what kind of weird agar recipe I could personally create, and it just hit me when I had both ingredients in hand. I have no other purpose for this, but only to see what happens. I thought cookies would be a better agar additive than beef...

Quote:

let us know how it turns out.




That is the whole purpose. To test FastFred's recipes againsed a "tried and true" recipe, and post the results. I will explain in greater detail when the time comes...


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Offlinekilgore_trout
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Re: FastFred's Media Cookbook [Re: ATWAR]
    #3064551 - 08/29/04 08:34 AM (19 years, 9 months ago)

definitely better than beef. bc i mean they have sugar and flour and what not and i mean i thought blood agar was for bacteria and what not mostly. i cant imagine using that shit for shrooms.


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Invisiblefastfred
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Re: FastFred's Media Cookbook [Re: kilgore_trout]
    #3067927 - 08/30/04 03:25 AM (19 years, 9 months ago)

I just posted an updated version of the cookbook! I updated it by adding an index, moving the sources to the end, and removing the chemical analysis of malt and yeast extract. I also added a few recipes including Karo, honey water, and dextrose tek.

What do you guys think? Better, worse, or indifferent?

I still plan to write an introduction and some general notes. Anything else I should add?


-FF

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Offlineragadinks
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Re: FastFred's Media Cookbook [Re: fastfred]
    #3068008 - 08/30/04 04:34 AM (19 years, 9 months ago)

I think it's better.
Maybe you could also add Una's Oatmeal Agar to the basic recipes which is also is a very good agar for certain species ?
And a cardboard agar recipe might also be interesting.
See this thread at mycotopia.


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-> It Is Not Important WHO Is Right But WHAT Is Right <-

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