Home | Community | Message Board


This site includes paid links. Please support our sponsors.


Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: Unfolding Nature Unfolding Nature: Being in the Implicate Order   PhytoExtractum Buy Bali Kratom Powder   Original Sensible Seeds Bulk Cannabis Seeds   OlympusMyco.com Olympus Myco Bulk Substrate   Left Coast Kratom Buy Kratom Extract   MagicBag.co All-In-One Bags That Don't Suck   Mushroom-Hut Grow Bags   Kraken Kratom Kratom Capsules for Sale   Bridgetown Botanicals Bridgetown Botanicals   Myyco.com Golden Teacher Liquid Culture For Sale   North Spore Bulk Substrate

Jump to first unread post Pages: 1 | 2 | 3 | 4  [ show all ]
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
FastFred's Media Cookbook (v0.97) Now 60 Recipes, Indexed! * 2
    #2996232 - 08/12/04 03:27 PM (19 years, 9 months ago)
Log in to view attachment

FastFred's Media Cookbook (v0.97)(8-30-2004)
--------------------------------------------------

by FastFred and ChesterKarma


...Index...
Section A. Classic Favorite Recipes
------------------------------------
Potato Dextrose Agar (PDA)(FDA M127)
Potato Dextrose Yeast (PDY)
Malt Extract Agar (3%)(MEA)(General Microbiology)(FDA M93)
Malt Extract Agar with Yeast (2%)(MEAY)
Sabouraud's Dextrose Broth and Agar (SabDex)(FDA M133)

Section B. Special Blends
--------------------------
"Karo Water" (Corn Syrup Broth)
"Dextrose Tek" Liquid Media (Corn Sugar Broth)
Honey Water (Mycotopia Honey Tek)
Malt-Yeast-Peptone Agar (McKenna's MYP)
Malt Yeast Peptone Agar (Stamets MYP)
Grey Cardboard (instead of agar)
Oatmeal Flake Agar
Moonflower's Rice Malt-Alfalfa-Brewer's Yeast Agar
MycoPsycho's Liquid Mycelial Culture Broth [Malt base]
Ragadinks Liquid Medium
Amaranth Soy Agar
EntheoGenesis No.442

Section C. Food Based Recipies and Variations
------------------------------------------------
Corn Meal Agar (CMA)
Cornmeal Dextrose Agar
Potato Flake Agar
Potato Starch Agar
Potato Flake Agar with Yeast
Potato-Carrot Agar
Barley Flour Malt Extract Agar
Barley Flour Modified Sabouraud's
Oatmeal Agar (OA)
Oatmeal Agar A
Oatmeal Agar B
V-8 Oatmeal Agar
V8 Medium
Bean Agar
Faba Bean Dextrose Agar (FDA)
Pea Agar
Cabbage Agar
Dr. Pollock's Modified [Dog Food] Agar

Section D. Other Media and Alternate Formulations
-----------------------------------------------------
Potato-Glucose Agar 1
Potato-Peptone Medium
Potato-Peptone-Yeast Agar (PPYA)
-
Malt Agar (MA)(FDA M185)[aka 2% MEA]
Difco Malt Extract Broth (FDA M94)
Malt Extract Agar for Yeasts and Molds (MEAYM)(FDA M182)
Malt Extract Peptone Agar
Raper & Thom MEA (RTMEA)
ISP 2 Medium (Malt, Yeast, Glucose)
-
Yeast Extract Agar (YEA)(FDA M181)
Yeast Glucose Agar
Glucose and Yeast Extract Agar
Glycerin Yeast Agar
-
Manure Tincture
Manure Agar
-
Starved Agar
Gelatin Agar (GA)(FDA M54)
Plate Count Agar (SMA)(aka Standard Methods Agar)(FDA M124)
Nutrient Agar (FDA M112)
Starch Agar (FDA M143)(Nutrient agar with starch)
Starch-Yeast Agar
1/5 Starch-Yeast Agar
Long-term Preservation Medium (FDA M85)
Peptone Meat Agar (Meat Water)

Section E. Common Solutions
------------------------------
Gentamicin Sulfate Solution (FDA M57)

Section F. Sources
-------------------
List of sources.


The whole thing doesn't seem to want to show up, so grab the text file from the top of the post.
What's your favorite media(s)?
You may choose many


Votes accepted from (08/12/04 12:00 AM) to (No end specified)
You must vote before you can view the results of this poll


Edited by fastfred (05/02/06 11:25 PM)

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: fastfred]
    #2996265 - 08/12/04 03:34 PM (19 years, 9 months ago)

I decided to post my list of common media recipes. I realize that there are probably similar guides out there. Please post any links, additional recipes, and corrections.

I plan to tidy it up a bit and add an index. Some of the entries are redundant, but I think that's helpful to see the similarities and slight differences. I also need to add some more recipes and find a nutritional analysis for potato infusion. I would like to add DFA, honeywater, and karo also, so if anyone has a favorite recipe or if there is already a consensus on the best karo/honeywater please let me know.


-FF

Extras: Filter Print Post Top
OfflineKillBill
Quick Learner
Registered: 07/02/04
Posts: 109
Last seen: 19 years, 8 months
Re: FastFred's Media Cookbook [Re: fastfred]
    #2996414 - 08/12/04 04:17 PM (19 years, 9 months ago)

Well i said Honey Water and Karo cause those are the only two i know how to do now. I can't find malt, i cant' find dextrose, i can find potatoes, but do i really feel like doing any (even minor) amounts of chemistry?
Teaspoon, dump two of whatever into water.. cook... Done.

I would like to try malt though, i hear its fastest (mycopsycho)

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #2996461 - 08/12/04 04:32 PM (19 years, 9 months ago)

I use a following mixture as a liquid medium and it works quite well:

16.5 g dextrose
1.5 g yeast
500 ml tap water

Here is a picture of mycelium growing in that broth:


Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #2996511 - 08/12/04 04:40 PM (19 years, 9 months ago)

Btw.: Have found a huge list of different agar media at dsmz:

http://www.dsmz.de/media/media.htm

Edited by ragadinks (08/12/04 04:46 PM)

Extras: Filter Print Post Top
Invisiblemycopsycho
Tit Inspector.
Male
Folding@home Statistics
Registered: 06/17/04
Posts: 3,712
Loc: Going Nowhere Fast
Re: FastFred's Media Cookbook [Re: fastfred]
    #2996548 - 08/12/04 04:46 PM (19 years, 9 months ago)

we all know what i like......


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ragadinks]
    #2996585 - 08/12/04 04:53 PM (19 years, 9 months ago)

Thanks for the link.  Most of the info in the cookbook comes from here:
FDA Bacteriological Analytical Manual(BAM) Media Index

I've tried to stick with simple medias that are known to work.

Come up with a title, description, and the source you want listed and I'll add it to the cookbook! :thumbup:


-FF

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #2996754 - 08/12/04 05:29 PM (19 years, 9 months ago)

Here is another good article from www.mushworld.com that examines the influence of different media on the growth of fungi:

Quote:

The morphology and growth rate of over 70 strains of 20 species of edible and medicinal mushrooms from the culture collection of N.G. Kholodny Institute of Botany, National Academy of Sciences of Ukraine were studied on 5 agar media. The optimal incubation time, temperature, and suitable media were determined for each culture. One natural and three synthetic liquid media with mineral and organic nitrogen sources were also tested. The effect of medium pH on the mycelial growth was determined by using phosphate buffers with pH values between 3.0 - 8.0. In each stage of inoculum preparation we used physiologically active mycelia. Factors affecting mycelial growth of some medicinal mushrooms in submerged culture were investigated. Inoculum size and composition and nutrient medium pH were important for maximising growth rate and biomass yield and minimising the duration of the lag phase of growth.



Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: KillBill]
    #2996783 - 08/12/04 05:33 PM (19 years, 9 months ago)

Quote:

KillBill said:
Well i said Honey Water and Karo cause those are the only two i know how to do now. I can't find malt, i cant' find dextrose, i can find potatoes, but do i really feel like doing any (even minor) amounts of chemistry?




Table sugar is sucrose. It is composed of two simpler sugars called glucose(dextrose) and fructose. Glucose is often called dextrose or corn sugar, and is made from cornstarch.

There isn't any chemistry involved in making PD. If you can cook sliced potatos (don't even have to peel them) for 30 min, then you can make PD broth.

Use corn sugar for the dextrose. If you can't find that, just use table sugar (sucrose).


-FF

Extras: Filter Print Post Top
Invisiblemycopsycho
Tit Inspector.
Male
Folding@home Statistics
Registered: 06/17/04
Posts: 3,712
Loc: Going Nowhere Fast
Re: FastFred's Media Cookbook [Re: fastfred]
    #2996870 - 08/12/04 05:49 PM (19 years, 9 months ago)

table sugar should be a last resort though.


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.

Extras: Filter Print Post Top
OfflineRaadt
nicht

Registered: 06/07/02
Posts: 2,107
Loc: azurescending
Last seen: 5 years, 1 month
Re: FastFred's Media Cookbook [Re: fastfred]
    #3000713 - 08/13/04 12:35 PM (19 years, 9 months ago)

You're killing me here.

Glucose - A monosaccharide sugar, C6H12O6

Dextrose - C6H12O6?H2O -- not a monosaccharide.


Very similar in structure, but not the same.

And while corn syrup may contain dextrose, glucose, and fructose. It is most certainly not a simple sugar.

They are interchangeable, sure. But saying they are the same thing is poor misinformation.


Very nice compilation of medias though. Thank you for the post.

Extras: Filter Print Post Top
OfflineSpudz76
Got mycelium on the brain...
Male

Registered: 02/08/04
Posts: 391
Last seen: 14 years, 2 months
Re: FastFred's Media Cookbook [Re: mycopsycho]
    #3000746 - 08/13/04 12:43 PM (19 years, 9 months ago)

Quote:

mycopsycho said:
we all know what i like......



Boobies!?!?  :eek:





oh, and malt extract too I guess. :smile:

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: Raadt]
    #3001303 - 08/13/04 02:43 PM (19 years, 9 months ago)

Quote:

Raadt said:
Glucose - A monosaccharide sugar, C6H12O6

Dextrose - C6H12O6?H2O -- not a monosaccharide.




Glucose and dextrose monohydrate are the same. There are hydrated and monohydrated forms of both. Check out sigma's website...

Dextrose, USP, anhydrous
Synonyms: D-(+)-Glucose, grape sugar, corn sugar
Molecular Formula: C6H12O6

Dextrose monohydrate, USP
Synonyms: D-(+)-Glucose monohydrate, glucose
Molecular Formula: C6H12O6?H2O

So... table sugar(sucrose) is a mixture of fructose and dextrose(glucose) monohydrate. Corn sugar is dextrose. Dextrose is assumed to be in it's nonhydrated form if not specified.


Quote:

Very similar in structure, but not the same.




I assume that the invivo conversion is trivial to the organism.


Quote:

And while corn syrup may contain dextrose, glucose, and fructose. It is most certainly not a simple sugar.




I don't know about corn syrup, but corn sugar is dextrose(glucose).


Quote:

Very nice compilation of medias though. Thank you for the post.




Thanks Raadt. I plan to HTMLize it at some point, after I've expanded it a bit.


-FF

Extras: Filter Print Post Top
Invisiblemycopsycho
Tit Inspector.
Male
Folding@home Statistics
Registered: 06/17/04
Posts: 3,712
Loc: Going Nowhere Fast
Re: FastFred's Media Cookbook [Re: Spudz76]
    #3001677 - 08/13/04 04:16 PM (19 years, 9 months ago)

Quote:

Spudz76 said:
Quote:

mycopsycho said:
we all know what i like......



Boobies!?!?  :eek:
oh, and malt extract too I guess. :smile:



yeah.... i like both of them.


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.

Extras: Filter Print Post Top
InvisiblePsyllyMe
LOST.......
Registered: 07/25/04
Posts: 848
Loc: Center of the Known Unive...
Re: FastFred's Media Cookbook [Re: mycopsycho]
    #3001732 - 08/13/04 04:39 PM (19 years, 9 months ago)

Malt n Boobs are great!

Wanna try my Sierra Nevada kill your spores tek?

Extras: Filter Print Post Top
Invisiblemycopsycho
Tit Inspector.
Male
Folding@home Statistics
Registered: 06/17/04
Posts: 3,712
Loc: Going Nowhere Fast
Re: FastFred's Media Cookbook [Re: PsyllyMe]
    #3001735 - 08/13/04 04:40 PM (19 years, 9 months ago)

lol..... no, my spores have a zest for life.


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.

Extras: Filter Print Post Top
InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: FastFred's Media Cookbook [Re: fastfred]
    #3004840 - 08/14/04 04:11 PM (19 years, 9 months ago)

Impressive copy&paste job :shake:

Maybe it would be nice to mention that half of the recipes you give are not intended for the cultivation of mushroom mycelia but for the cultivation of bacteria. Mushroom mycelia will grow very poorly on these media.

For fungi just stick to malt extraxt agar, potato dextrose agar and oatmeal agar and forget about exotic recipes like PCA or Sabouraud


--------------------
www.911blogger.com

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: Una]
    #3009482 - 08/16/04 02:56 PM (19 years, 9 months ago)

Thanks Una. It is indeed a copy&paste job. My intention was to save people some time by collecting this info together. It's much easier than wading through 100s of media types. I'm hoping it will grow into more than just a cut&paste collection as people add their own comments, recipes, and ideas.


>Maybe it would be nice to mention that half of the recipes you give are not intended for the cultivation of mushroom mycelia but for the cultivation of bacteria.<

I just picked out all the media types that are both simple to make and will grow mycelium. Growth speed is not a major factor in petri dish cultivation. Sometimes it's useful to change the media for your specific needs. It's also good to change up the media to keep your mycelium vigorous. Just switching up PDA and MEA is kind of limited.


>For fungi just stick to malt extract agar, potato dextrose agar and oatmeal agar and forget about exotic recipes like PCA or Sabouraud<

From: PML Microbiologicals
"Sabouraud Dextrose (SabDex) Agar is used for the isolation, cultivation, and maintenance of saprophytic and pathogenic yeasts and fungi.

SabDex Agar is an excellent substitute for Malt or Potato Dextrose Agar, when used by mushroom cultivators to propagate mushroom mycelium."

Uses include testing homes, basements, schools, businesses, and vehicles for toxic mold.

Sabouraud Dextrose Agar was described by Sabouraud in 1892 and was used for the identification of fungi based on their morphological characteristics. Sabouraud Dextrose Agar is a standard medium used to support the growth of yeasts and molds. It supplies peptone as the protein source and dextrose as the carbohydrate source for nourishment. Bacterial suppression occurs due to the low pH. This media is especially suited for the primary isolation of fungi from normally sterile sites such as cerebrospinal fluid (CSF).

Later, Emmons modified the medium by decreasing the dextrose content and adjusting the pH closer to the neutral range. This modification enhances sporulation and is particularly useful for the subculture of fungi that so not develop fruiting structures on other media, and so is useful in their identification. It also serves as a good holding medium for stock cultures.

Source: PML Microbiologicals


If you've tried any of the media types in my list an find that they don't work, please post a message or PM me and I'll change the guide.

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: FastFred's Media Cookbook [Re: fastfred]
    #3014550 - 08/17/04 04:04 PM (19 years, 9 months ago)



--------------------
To give is to live...


Edited by ATWAR (08/17/04 04:15 PM)

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: ATWAR]
    #3022176 - 08/19/04 05:16 AM (19 years, 9 months ago)

It strange that a lot of professional culture/mycology institutes recommend a much shorter sterilization time then it says in the link you provided:
Quote:

All media have a near-neutral pH and should be sterilized for twenty to thirty minutes at 15 psi pressure.




Here it says 20-30 minutes whereas some site recommend 10-15 minutes:
Here 10 minutes are recommended and here 10 - 15 minutes are recommended and here at the dmsz also 10 minutes are recommended.
I would rather stay with the 10-15 minutes, what do you think ?

Extras: Filter Print Post Top
InvisibleSpeeker

Registered: 02/11/04
Posts: 894
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3022292 - 08/19/04 06:13 AM (19 years, 9 months ago)

And Mr.Paul Stamets says in GGMM:
Quote:

Often the medium upon which a culture is grown becomes the source of contamination. Insufficient sterilization is usually the cause. Standard sterilization time for most liquid media is only 15-20 minutes at 15 psi or 250 F. (121 C.). However, this exposure time is far too brief for many of the endospore-forming bacteria prevalent in the additives currently employed by cultivators. I recommend at least 40 minutes @ 15 psi for malt extract or potato dextrose agars.




:wink:

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: Speeker]
    #3022356 - 08/19/04 06:54 AM (19 years, 9 months ago)

Good point :wink:.

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3023678 - 08/19/04 01:32 PM (19 years, 9 months ago)

What is Stamets talking about?

"However, this exposure time is far too brief for many of the endospore-forming bacteria prevalent in the additives currently employed by cultivators."

What additives? 15-20 Minutes is the standard time... If you are using standard media the standard time that has worked for decades for microbiologists is fine. If you use a non-standard sterilization time, then you're not making standard media. You're making overcooked, non-standard media.

Cooking your malt as long as Stamets suggests may caramelize it. I highly doubt that the FDA and the Bacteriological Analytical Manual, 8th Edition, Revision A, 1998 don't know how to sterilize media. They recommend 15 min @ 121 deg C. That is the de facto standard. Anything more is wasteful overcooking. If you want to be safe go for 20 min.

I realize that dissing Stamets is a sure way to get flamed, but in this case I think Stamets is out of his head and overstepping his bounds. He may have had fine success with overcooking his media, but that doesn't provide any reason to dispute the fact that 15-20 minutes will sterilize PDA, PDY, and MEA just fine and is the undisputed and fully standardized way to sterilize media.


-FF

Edited by fastfred (08/19/04 03:56 PM)

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #3023728 - 08/19/04 01:44 PM (19 years, 9 months ago)

Hmm, I think it depend on what kind of additives you are adding.
Stamets uses soil, sawdust and stuff like this in order to keep the mycelium "busy".
Maybe that's why he uses that long sterilization times?
I myself have reduced the sterilization times to 15 minutes and had no problems so far even when using a bit of sawdust as additive and not adding peroxide at all.

Extras: Filter Print Post Top
OfflineRaadt
nicht

Registered: 06/07/02
Posts: 2,107
Loc: azurescending
Last seen: 5 years, 1 month
Re: FastFred's Media Cookbook [Re: fastfred]
    #3023751 - 08/19/04 01:48 PM (19 years, 9 months ago)

FF:

You are wrong. Longer exposure to the same temperature will not cause anything to caramelize any differently. Caramelization occurs at ~320f, 250 is 250 is 250 is 250. Otherwise, wouldn't your sugars in your grains caramelize as well?

From now on please refer to a reference if you are posting something such as media will caramelize if you cook it longer than 20 minutes. Because you're looking more and more a fool, to me.

I have cooked media (which i had ground grain in) for 45 minutes. And I have never had any such problem.

Extras: Filter Print Post Top
OfflineAnnoA
Experimenter
 User Gallery

Folding@home Statistics
Registered: 06/17/99
Posts: 24,166
Loc: my room
Last seen: 23 days, 22 hours
Re: FastFred's Media Cookbook [Re: Raadt]
    #3023805 - 08/19/04 02:01 PM (19 years, 9 months ago)

http://www.exploratorium.edu/cooking/candy/caramels-story.html

"Caramels are the chewy candies you are familiar with. They?re made by cooking sugar, cream, corn syrup, and butter to 245? F. Their brown color comes from a reaction between the sugar and the protein in the cream. This reaction is called the Maillard reaction, after the French scientist who discovered it. The rich brown color of toasted nuts and barbecued meats also comes from the Maillard reaction.

Put simply, the Maillard reaction occurs when part of the sugar molecule (the aldehyde group, if you must know) reacts with the nitrogen part of the protein molecule (an amino group). The resulting series of reactions is not well understood even by food scientists, but it leads to the brown color and many flavorful compounds that are yet to be identified."

So the term used in mushroom circles "caramelisation" is actually incorrect and should be "Maillard reaction".

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: Anno]
    #3024141 - 08/19/04 03:17 PM (19 years, 9 months ago)

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.

Where is your citation that carmelization only occurs at +320F? Everything I've read suggests that carmelization is a problem when making media. None of these people are using super high sterilization temps and they go to great lengths to prevent carmelization.

If you're going to call me a fool, you better have some good information, otherwise you end up looking not only foolish, but arrogant.

It also makes sense that different sugars carmelize differently and that other ingredients in the media affect carmelization.

"YPD will turn a rich brown color on autoclaving due to carmelization of the dextrose." -Forrest Spencer, Positional Cloning Course, Cold Spring Harbor 1998

"Carmelization - Cane sugar deteriorates in heated conditions to form a colored breakdown known as carmelization." -Sugar Confections How candies are made by The Nut Factory

"If Dextrose is added before autoclaving, the sterilized medium will be darker due to carmelization of the sugar. Otherwise, it is often useful to have sterile YP medium to which different carbon sources (e.g., Dextrose, Galactose, Glycerol) can be added." -bioProtocol a Bio online site

"Sorbose (used to induce colonial growth) caramelizes when autoclaved if medium N or another medium containing NH4 is used. A brown breakdown product is produced. This does not usually seem to affect colonialization. But carmelization can be avoided by autoclaving a sorbose-glucose-fructose 2OX solution separately from the medium and combining after sterilization (D.E.A. Catcheside), or more simply by substituting Synthetic Cross medium (which is NH4-free) for Medium N (T. Legerton)." -David D. Perkins - Hints and precautions for the care, feeding and breeding of Neurospora

"Carmelization: brown color due to breakdown of sugars at high temperatures" -Carbohydrates Chapter 3

"To reducecarmelization, the L-sorbose was autoclaved separately fromthe other ingredients." -USE OF RAPD, ENZYME ACTIVITY STAINING, AND COLONY SIZE TO DIFFERENTIATE PHYTOPATHOGENIC FUZARIUM OXYSPORUM ISOLATES FROM IRAN


-FF

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: Anno]
    #3024212 - 08/19/04 03:30 PM (19 years, 9 months ago)

Here s a good and detailed paper about the "Maillard reaction" and it seems that the thing are not so easy:
The reaction can take place below 245? F (118,3? C) and depends very much on the kind of sugar, water content, PH, duration and temperature of the reaction and available oxygen.
So it very probable that heating at 121? C for some time starts the "Maillard reaction". The question is if the caramelization is strong enough to influence the growth of the mycelium.
Some agar recipes recommend a short sterilization time and the addition of activated charcoal in order to avoid chemical byproducts of the sterilization process that are harmful to the mycelium.

The following excerpt is from this paper:

Quote:

1. Murashige & Skoog medium (MS) (Table 5) was supplied with antibiotics (50 ppm streptomycin and ampicillin, 5 ppm benomyl) and then sterile filtered (0.2 um Sartorius cellulose acetate filter). MS was then mixed with an autoclaved agar solution (final agar concentration 1.2%) with 0.05% activated charcoal (to adsorb toxic compounds in autoclaved agar), before addition to 9-cm petri dishes.




The question is if the mentioned toxic compounds are a result of the Maillard reaction or of something else or of a lot of different reactions?

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3024246 - 08/19/04 03:42 PM (19 years, 9 months ago)

It is my understanding that caramelization is not a scientific term and does not refer to a specific chemical reaction. Instead it refers to multiple reactions with a wide variety of different sugars.

The dictionary says:
caramelization - To convert or be converted into caramel.
caramel - 1. Burnt sugar, used for coloring and sweetening foods.
2. A moderate yellow brown.

Using this definition "caramelization" refers to the heating of sugar which results in darkening or colorization.

That definition fits in line with the usage of the term in relation to media. If it has sugar in it and you cooked it and it darkened... Then it can be said to have undergone some degree of caramelization.


-FF

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: Raadt]
    #3024347 - 08/19/04 04:01 PM (19 years, 9 months ago)

Quote:

You are wrong. Longer exposure to the same temperature will not cause anything to caramelize any differently. Caramelization occurs at ~320f, 250 is 250 is 250 is 250. Otherwise, wouldn't your sugars in your grains caramelize as well?



Yes the do and it seems as if you were wrong ...

Quote:

From now on please refer to a reference if you are posting something such as media will caramelize if you cook it longer than 20 minutes. Because you're looking more and more a fool, to me.



In this case you should come up with reference, shouldn't you ?
Otherwise someone else might look like a fool ...

Quote:

I have cooked media (which i had ground grain in) for 45 minutes. And I have never had any such problem.



As said in another posting: caramelization depends on a lot of factors and not all strains are so sensitive to it.
But you certainly have a more or less small amount of caramelized sugars and I think they are at least lost as a source of nutrition for the mycelium if not harmful.

The point at the whole discussion with you is, that you are quite arrogant to other people and at the same time you do the same things you accuse them to do: You do not back up your statement with profound knowledge.
In this thread you say for example that chanterelles cannot be cultivated - they can ...

I mean the point is that we all make mistake and are sometimes wrong in our statements like you, me and fastred.
But if we would not make mistakes we would not have a discussion going on that might be instructive for some or all of us.
But the discussions in which you take place do not have the goal to gain some information but to show your superiority over the others and that is usually a sign of low self-esteem.

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3024469 - 08/19/04 04:24 PM (19 years, 9 months ago)

Thanks for the great link ragadinks! This should go a long way towards settling the Maillard reaction vs caramelization debate.

The Maillard Reaction: Application to Confectionery Products by C.G.A. Davies & T.P. Labuza

From that paper:
----------------
The Maillard reaction is one of four nonenzymatic browning reactions which occur in
food. The other three are:
1) the degradation of ascorbic acid,
2) lipid peroxidation
3) sugar-sugar caramelization.
[...]
At high temperatures (> 80 oC[176F]) sugar-sugar interactions or the caramelization reaction occurs. This is a complex series of reactions but many of the intermediate flavor compounds and products are similar to those observed for the Maillard reaction.
----------------

So it looks like Raadt is completely wrong... I don't mean to gloat, and I certainly don't want to hurt Raadt's self-esteem... But turnabout is fair play Raadt. Let this be a lesson next time you think about calling someone a fool.


Who is the fool now Raadt?


-FF

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: FastFred's Media Cookbook [Re: fastfred]
    #3024530 - 08/19/04 04:46 PM (19 years, 9 months ago)

Quote:

fastfred said:
Who is the fool now Raadt?






I hate to see stuff like this, it kills a good converstation. Let's try to keep it professional, ok?

We are all human, and we all make mistakes. Let us be who we are. (used to be my old sig)

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: ATWAR]
    #3024542 - 08/19/04 04:50 PM (19 years, 9 months ago)

I agree - that is going to lead to nowhere ...

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3024583 - 08/19/04 05:06 PM (19 years, 9 months ago)

It's all well and good to talk about turning the other cheek... But where were you guys when Raadt was calling me a fool?

As I said earlier turnabout is fair play. If Raadt had presented his unsupported ideas about caramelization in a less arrogant way, presented them as conjecture rather than fact, or simply not called me a fool we would not be having this problem.

I think Kid Rock says it best:

A lot of people poke fun and that's alright
But when I start pokin' back they get all uptight
Huh!
You can't cap with the master, son
So sit your ass down before I blast ya one



-FF

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: FastFred's Media Cookbook [Re: fastfred]
    #3024621 - 08/19/04 05:19 PM (19 years, 9 months ago)

Have you ever heard two wrongs do not make a right?
Both you and Raadt were wrong for making personal attacks agained one another. We would not be having this problem if you just let it go when the info was shown to be wrong. If you had just left it, there would be no continuation of the problem, instead you added to it... Like I said, things like this kill a thread...

Ones actions speak very much about ones character...

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #3024662 - 08/19/04 05:29 PM (19 years, 9 months ago)

I agree with ATWAR.
Raadt was wrong and arrogant but now it looks like as if you wanted to take his place ...

For me this place should not be a battlefield were the other one wins and the other one looses but a place where you can meet nice people and learn something about mushrooms.
So, and that's the last posting I will loose about shitty that topic anymore ...

Extras: Filter Print Post Top
Offlinefugu
Pooh-Bah

Registered: 01/06/03
Posts: 2,223
Loc: istanbul
Last seen: 13 years, 11 months
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3024814 - 08/19/04 05:59 PM (19 years, 9 months ago)

  :thumbup:


--------------------
mushroom culture history making ...Mr. Allan is the best .....

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: fugu]
    #3025057 - 08/19/04 06:42 PM (19 years, 9 months ago)

5 posts have have now been wasted bitching and moaning about 1 line of text which was 100% fair play.

Two wrongs don't make a right... and 6 wrongs is even worse.

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.

If you can't handle a spirited discussion which sometimes escalates because of some turnabout, then I suggest you guys find one of those "kid-safe" sites where even the most minor putdown is deleted before it ever gets posted.

If that doesn't appeal to you, then grow up and quit your worthless bitching.

-FF

P.S. Raadt may have been wrong and even insulted the correct posters, but at least he's grown up enough to take his lumps and not whine and cry about it like a bunch of babies.

Extras: Filter Print Post Top
Anonymous

Re: FastFred's Media Cookbook [Re: fastfred]
    #3025271 - 08/19/04 07:18 PM (19 years, 9 months ago)

ROFLMAO!

I just joined and your media cookbook has been a big help for me fastfred.

I think you are fully justified in calling Raadt a fool. He'll be much more carefull about talking jive and dissin' others in the future I bet.

As for the rest of the people complaining about you, what a bunch of whiners. It's stupid how many people think it's cool to bitch about bitching. Makes them look like a bunch of bitches. I hope the rest of the site is more constructive and less bitchy.

They'll probably write a dozen posts to bitch about this post! Oh, well I have what I need anyway. Go ahead and bitch on ATWAR, ragadinks, and the rest of the bitches.

Extras: Filter Print Post Top
Anonymous

Re: FastFred's Media Cookbook [Re: ]
    #3025302 - 08/19/04 07:24 PM (19 years, 9 months ago)

Damn, with all that bitching I forgot to ask my question. What is the best media for a noob like me? I can't get anything fancy.

Also sorry if I upset any of the grand poo-bitches of this board with my anti-bitch bitching.

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: FastFred's Media Cookbook [Re: ]
    #3025865 - 08/19/04 09:12 PM (19 years, 9 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...


--------------------
To give is to live...


Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ATWAR]
    #3025935 - 08/19/04 09:27 PM (19 years, 9 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

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: FastFred's Media Cookbook [Re: fastfred]
    #3026039 - 08/19/04 09:54 PM (19 years, 9 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)...

Extras: Filter Print Post Top
InvisibleSpeeker

Registered: 02/11/04
Posts: 894
Re: FastFred's Media Cookbook [Re: fastfred]
    #3026972 - 08/20/04 01:55 AM (19 years, 9 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)

Extras: Filter Print Post Top
Offlineliveby
Wasted For Time
 User Gallery

Registered: 06/15/04
Posts: 1,511
Last seen: 13 years, 1 month
Re: FastFred's Media Cookbook [Re: fastfred]
    #3027002 - 08/20/04 02:10 AM (19 years, 9 months ago)

I did pasta and rice!

Extras: Filter Print Post Top
InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: FastFred's Media Cookbook [Re: fastfred]
    #3031793 - 08/21/04 07:27 AM (19 years, 9 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.


--------------------
www.911blogger.com

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: Una]
    #3036789 - 08/22/04 04:37 PM (19 years, 9 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

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #3036846 - 08/22/04 04:54 PM (19 years, 9 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 ?

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3036880 - 08/22/04 05:04 PM (19 years, 9 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

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
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...

Extras: Filter Print Post Top
InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
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?


--------------------
www.911blogger.com

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

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
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.



Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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

Extras: Filter Print Post Top
InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
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.


--------------------
www.911blogger.com

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
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


--------------------
To give is to live...


Extras: Filter Print Post Top
Offlinekilgore_trout
Stranger
Registered: 10/17/03
Posts: 1,607
Last seen: 15 years, 10 months
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.


--------------------
"I didnt fight a secret war in nicaragua so you could walk these streets of freedom bad-mouthing lady america in your damn mirrored sunglasses."

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
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...


--------------------
To give is to live...


Extras: Filter Print Post Top
Offlinekilgore_trout
Stranger
Registered: 10/17/03
Posts: 1,607
Last seen: 15 years, 10 months
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.


--------------------
"I didnt fight a secret war in nicaragua so you could walk these streets of freedom bad-mouthing lady america in your damn mirrored sunglasses."

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
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.


--------------------
-> Errors Are A Great Source Of Knowledge <-
-> It Is Not Important WHO Is Right But WHAT Is Right <-

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: ragadinks]
    #3068022 - 08/30/04 04:55 AM (19 years, 9 months ago)

Done... Thanks for the input.


-FF

Extras: Filter Print Post Top
Invisiblemycopsycho
Tit Inspector.
Male
Folding@home Statistics
Registered: 06/17/04
Posts: 3,712
Loc: Going Nowhere Fast
Re: FastFred's Media Cookbook [Re: fastfred]
    #3068730 - 08/30/04 10:57 AM (19 years, 9 months ago)

i'm loving the update fred! good job and keep it coming!


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #3069751 - 08/30/04 03:18 PM (19 years, 9 months ago)

Do not know wether it's helpful but I have found recipes for antibiotics solution here.

Extras: Filter Print Post Top
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: FastFred's Media Cookbook [Re: fastfred]
    #3159164 - 09/21/04 08:27 AM (19 years, 8 months ago)

Ok, here are the results from the media I tested. As predicted, the selected media performed very poorly againsed the control MYPA (recipe from GGMM, 2% Malt). The most striking difference in performance was experienced with Ganoderma lucidum. There was a huge difference in speed of colonization between the control plate and the ?questionable? media recipes with the Reishi:



And of course, I had to test the most commonly grown mushroom here, Psilocybe cubensis. The mycelium of Ps. C. grew extremely fine, and not quite as quickly as the control:



Then there was Pleurotus ostreatus, a perfect mushroom for doing experiments with as its growth characteristics are well known and predictable. It is interesting to note that only the control and the Gelatin Agar colonized. The other two mediums failed to grow mycelium further than ?? away from the wedge?



And I also tested Trametes versicolor. It is my experience that the Turkey Tail has very aggressive mycelia, and did indeed colonize the test plates at exactly the same rate as the control. However, the mycelial mat was not nearly as thick or vigorous. The other two medias missing (forgot to take pictures) performed identically to the Nutrient Agar pictured below:




I made up a 2nd batch using Coprinus comatus and Hericium erinaceus, but that was about a week ago. I got tired of waiting so I posted these earlier results... I also left out my cookies and milk agar, which performed slightly better than these test plates (although I got pins on the ostreatus CNM plate). Anyway, there you have it. All media is not created equal, and does not perform equally well in the end. If you want to experiment, go ahead and try recipes not designed for mushroom mycelia. But, if you want to grow quick and healthy mycelium, stick to the tried and true:

Una's proven agar nutrient media recipes.

Shroomery FAQ media recipes.

Shroomery Cultivation Guide media recipes.


--------------------
To give is to live...


Extras: Filter Print Post Top
Offlinehyphae
born to grow
 User Gallery

Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 13 years, 1 month
Trusted Cultivator
Re: FastFred's Media Cookbook [Re: ATWAR]
    #3160187 - 09/21/04 02:12 PM (19 years, 8 months ago)

So anyone heard from FF lately? Last thing I saw was some replying to FF simply saying "DEAD" an haven't heard a word since?

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: fastfred]
    #3212633 - 10/04/04 02:48 AM (19 years, 8 months ago)

I'm still around... Not dead yet... Who said I was dead? That's not a very nice thing to say.

I've gotten a new job, which is why I haven't posted lately. I've been busting my ass to pay the bills and whatnot.

I'd like to thank ATWAR for doing these experiments. Great job with the photos. I'll be revising the guide at some point to make clear that these medias perform poorly. Sorry for not getting back to you, but I got too busy.

I had expected that a several media types would not perform very well. I expected them to perform slightly better than they did though. Some of my imagined uses for some of the poorly performing medias seem much less usefull now that I've thought about them more.

I'll be doing some experiments relating to screening for auxotrophs in the comming months (hopefully), so if any media proves usefull in that respect, I'll add it to the guide.


-FF

Extras: Filter Print Post Top
Offlineragadinks
MrBeatle
 User Gallery

Registered: 10/20/03
Posts: 1,298
Last seen: 8 months, 1 day
Re: FastFred's Media Cookbook [Re: fastfred]
    #3212695 - 10/04/04 03:52 AM (19 years, 8 months ago)

> I'll be doing some experiments relating to screening for auxotrophs in the comming months (hopefully), so if any media proves usefull in that respect, I'll add it to the guide.
What does 'auxotrophs' mean ?

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: FastFred's Media Cookbook [Re: fastfred]
    #3215064 - 10/04/04 05:44 PM (19 years, 8 months ago)

An auxotroph is a mutant that can't grow in simple media. Usually it is a result of a mutation that knocks out a metabolism pathway. The result is that an auxotroph is unable to synthesize a certain amino acid, so it must be present in the substrate or it will be unable to grow.

They are mainly useful for mating experiments. If you have two auxotrophs of different strains they will likely be deficient in different areas, hence any fusion of the two will result in an organism that is nutritionally competent. It's a way to screen for successful matings and/or successful completion of the parasexual cycle.


-FF

Extras: Filter Print Post Top
Offlinehyphae
born to grow
 User Gallery

Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 13 years, 1 month
Trusted Cultivator
Re: FastFred's Media Cookbook [Re: fastfred]
    #3215255 - 10/04/04 06:30 PM (19 years, 8 months ago)

Good to hear your still around FF :wink:


--------------------
Getting the most out of your casings!, A pinning strategy.
Oyster Shell "Flour" $2 for 1lb. a hell of a deal :wink:
Not what is overlay but rather what overlay is
Gas Exchange vs. FAE

"We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"

Extras: Filter Print Post Top
OfflineCiv
Pinning
Male User Gallery

Registered: 10/14/04
Posts: 2,537
Loc: California
Last seen: 10 months, 29 days
Re: FastFred's Media Cookbook (v0.97) Now 60 Recipes, Indexed! [Re: fastfred]
    #3561929 - 12/30/04 03:07 PM (19 years, 5 months ago)

I have a question about the Oatmeal Flake Agar.

Quote:

Oatmeal Flake Agar
-------------------
75 g Oatmeal flakes
20-25 g Agar
1 L water

Stir for 5-10 minutes then filter out the larger particles by pouring it through some mesh, save the broth. [Then add the agar]





This is done post-PC correct? And do I grind up the oatmeal at all? Thanks.


--------------------
"...Gal's seem to hate the thought of blending chicken shit in a blender.
So, wash it well afterwards & DON'T tell them..."  -Agar

Extras: Filter Print Post Top
Offlinejeff01
cultivator
Registered: 12/17/04
Posts: 44
Last seen: 19 years, 5 months
Re: FastFred's Media Cookbook (v0.97) Now 60 Recipes, Indexed! [Re: Civ]
    #3563439 - 12/30/04 10:43 PM (19 years, 5 months ago)

great suff, thanks!


--------------------
"impossible to walk in this muck"

Extras: Filter Print Post Top
OfflineSnarp
Stranger
Registered: 01/28/05
Posts: 22
Last seen: 18 years, 11 months
Re: FastFred's Media Cookbook (v0.97) Now 60 Recipes, Indexed! [Re: jeff01]
    #3699877 - 01/29/05 08:09 PM (19 years, 4 months ago)

Civ: no and no.

FastFred: thanks. Too bad the rest of this thread was such a waste of time. Can't say I'm impressed by the maturity levels around here.

--Snarp

Extras: Filter Print Post Top
InvisibleIntelligentMind
Indigo Child
Registered: 01/26/05
Posts: 73
Re: FastFred's Media Cookbook [Re: fastfred]
    #3802575 - 02/19/05 03:51 AM (19 years, 3 months ago)

Happen to have that invitro agar recipe that Una uses for their salvia divinorum?! Now that, i'd like to know...

Extras: Filter Print Post Top
OfflineAnnoA
Experimenter
 User Gallery

Folding@home Statistics
Registered: 06/17/99
Posts: 24,166
Loc: my room
Last seen: 23 days, 22 hours
Re: FastFred's Media Cookbook [Re: IntelligentMind]
    #3802579 - 02/19/05 04:03 AM (19 years, 3 months ago)

The recipes for plant media differ very much from what you would use for mushrooms.

An excellent book on this topic is  Plants from Test Tubes : An Introduction to Micropropagation

Extras: Filter Print Post Top
InvisibleIntelligentMind
Indigo Child
Registered: 01/26/05
Posts: 73
Re: FastFred's Media Cookbook [Re: Anno]
    #3802604 - 02/19/05 04:32 AM (19 years, 3 months ago)

Thanks a lot, i'll be sure to check that book out.

Extras: Filter Print Post Top
OfflineAnnoA
Experimenter
 User Gallery

Folding@home Statistics
Registered: 06/17/99
Posts: 24,166
Loc: my room
Last seen: 23 days, 22 hours
Re: FastFred's Media Cookbook [Re: IntelligentMind]
    #3802637 - 02/19/05 05:11 AM (19 years, 3 months ago)


Extras: Filter Print Post Top
InvisibleIntelligentMind
Indigo Child
Registered: 01/26/05
Posts: 73
Re: FastFred's Media Cookbook [Re: Anno]
    #3805757 - 02/20/05 01:56 AM (19 years, 3 months ago)

*bows down to you* Good shit man! Thank you! Anno, you need any salvia cuttings? I'll send you one as a thank you. Peace man..

Extras: Filter Print Post Top
OfflineMycoJunkie
Psilanthropist

Registered: 11/04/04
Posts: 963
Loc: .4merica
Last seen: 18 years, 4 months
Re: FastFred's Media Cookbook [Re: IntelligentMind]
    #4069426 - 04/18/05 05:04 PM (19 years, 1 month ago)

bump


--------------------
:cussing::whip:

Extras: Filter Print Post Top
Jump to top Pages: 1 | 2 | 3 | 4  [ show all ]

Shop: Unfolding Nature Unfolding Nature: Being in the Implicate Order   PhytoExtractum Buy Bali Kratom Powder   Original Sensible Seeds Bulk Cannabis Seeds   OlympusMyco.com Olympus Myco Bulk Substrate   Left Coast Kratom Buy Kratom Extract   MagicBag.co All-In-One Bags That Don't Suck   Mushroom-Hut Grow Bags   Kraken Kratom Kratom Capsules for Sale   Bridgetown Botanicals Bridgetown Botanicals   Myyco.com Golden Teacher Liquid Culture For Sale   North Spore Bulk Substrate


Similar ThreadsPosterViewsRepliesLast post
* Potato Honey Agar frankz00 16,144 16 01/08/19 10:58 AM
by bodhisatta
* various liquid culture media Mindzpore 3,212 16 05/03/06 06:30 AM
by fastfred
* Some proven agar nutrient media recipes Una 12,219 11 10/03/04 01:15 PM
by YSr
* henderson hasselbach ph of media rungi 1,040 3 10/05/06 01:00 AM
by fastfred
* Does anyone use brown sugar as liquid media? Why not? forgieboy 3,062 14 09/05/04 03:46 PM
by Mycena
* PH in petri dishes? YSr 1,416 5 08/14/04 04:19 PM
by Una
* Re: Hello, can I sterilize media filled glass petris in the microwave? Anonymous 1,974 9 02/08/00 07:35 AM
by Anonymous
* media carmelization problem, cooker sterilization commersh 867 1 03/13/04 04:07 AM
by Joshua

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: RogerRabbit, Pastywhyte, bodhisatta
22,610 topic views. 0 members, 4 guests and 2 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2024 Mind Media. Some rights reserved.

Generated in 0.064 seconds spending 0.027 seconds on 17 queries.