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OfflineHumboldtHort
newbie
Registered: 02/13/02
Posts: 40
Last seen: 12 years, 8 months
Agar Advantage?
    #577474 - 03/13/02 01:14 AM (14 years, 8 months ago)

I was wondering about the advantages to agar? I was thinking about this and was curious if someone could help me out? When innoculating a jar with a syringe, you're introducing many spores to each other. Many dikaryotic mycelia arise from the joining of two capatible monokaryotic mycelium, right? Do the dikaryotic mycelia of the same species join with each other or not? If not, with agar, aren't you basically isolating a single dikaryotic mycelium with rhizomorps? How exactly does this work? Do you get one single network in a jar, or many with a syringe innoculation? Any light shed on this would be appreciated


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A gram is better than a damn


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: HumboldtHort]
    #577558 - 03/13/02 02:57 AM (14 years, 8 months ago)

an average PF style jar probably has 10+ different substrains colonising it, some of which will be sterile. The advantage of agar inocculation is that your isolated strain has the entire cake/jar of grain/binfull of straw to itself, rather than a small patch that it managed to colonise before the rest of the substrate got colonised by competing strains. Thinking about it, any bulk growing tech is a bit of a waste of substrate if it is spawned with a multispore grain jar, as only the substrains near the top of the casing will get the chance to fruit properly, and the all the underlying substrains do is effectively deprive the fruiting ones of substrate.


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hmmm........


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Offlinekarlfinn
member
Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
Last seen: 14 years, 4 months
Re: Agar Advantage? [Re: Champion des Champignons]
    #577671 - 03/13/02 08:59 AM (14 years, 8 months ago)

I was wondering....say...spore syrg. iinnocs. grain jar...they fight it out...
jusing a spoon of jar one to a new grain jar...gives the strain a better
full run into substrates...I know strain selects from agars are best, but
damn if that spore syrg is not so simple and clean..
spore injects to liquid probably the same fight...??
using this to innoc. grain jars....smaller competing populations....then
the grain to substrate.....could always islate one more time in jars...
not as slow as it may seem....all stronger in the end..


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: karlfinn]
    #577696 - 03/13/02 09:56 AM (14 years, 8 months ago)

yes, but there's no way of knowing if the most aggressive strain produces the best, if any, fruits.
The best thing to do is to fruit a multispore innoculation then clone the best fruiting strain. Even then there's no guarantee that the strain that produces the best fruit has a good colonising rate etc.
It takes a lot of trial and error before you come up with your ?ber strain.
Which is why we're not all master cultivators already:D 


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hmmm........


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InvisibleSwami
Eggshell Walker

Registered: 01/19/00
Posts: 15,413
Loc: In the hen house
Re: Agar Advantage? [Re: Champion des Champignons]
    #577747 - 03/13/02 11:18 AM (14 years, 8 months ago)

...and the all the underlying substrains do is effectively deprive the fruiting ones of substrate.

Is this your opinion or do you have references to any articles?


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The proof is in the pudding.


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Offlinekarlfinn
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Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
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Re: Agar Advantage? [Re: Champion des Champignons]
    #577947 - 03/13/02 04:40 PM (14 years, 8 months ago)

thanx for the info. and yup...nothing like a master culture...and clones...
I will go with fruit it , pick the best for r-d .....isloate that strain and...well pratice..practice....
otherwise...back to the agar plates ..I have an Earle and Amazonian from Rain Magic from 1988
no longer go by the name..but still in the biz ....
you know..those cotton swabs in the tube....found stored away...trying to rehydrate a while before plating
them out. pretty old spore but always kept cool, dry, sealed..we shall see. if so I shall see ya'll in the exchange
hopefully ...wishful thinking...and a challenge...


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OfflineHumboldtHort
newbie
Registered: 02/13/02
Posts: 40
Last seen: 12 years, 8 months
Re: Agar Advantage? [Re: Champion des Champignons]
    #578022 - 03/13/02 06:52 PM (14 years, 8 months ago)

C d C, do you have a master culture? Do you find that isolated mycelium cultures have a significantly higher yield than cultures with many mycelia in them? I can't currently undertake this task, but it sounds like something to look forward to. It seems that culturing mushrooms can be quite an artform.


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A gram is better than a damn


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: Swami]
    #578178 - 03/13/02 10:20 PM (14 years, 8 months ago)

Just an opinion, but it makes sense that substrate colonised by a competetor strain will prevent whichever strains that get a chance to fruit from using that bit of substrate.


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hmmm........


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: HumboldtHort]
    #578192 - 03/13/02 10:29 PM (14 years, 8 months ago)

I've just got into the whole agar/cloning business lately and have yet to get to the fruiting stage, but it makes sense to me that a substrain that has all the resources of a 10lb bin of straw at its disposal will be able to take advantage of it better than, say 10 different substrains battling it out amongst themselves.


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hmmm........


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OfflineHumboldtHort
newbie
Registered: 02/13/02
Posts: 40
Last seen: 12 years, 8 months
Re: Agar Advantage? [Re: Champion des Champignons]
    #578250 - 03/13/02 11:55 PM (14 years, 8 months ago)

Is this the route experienced cultivators usually take? I was looking for some answers on this in TMC, but didn't find any specific reasoning behind isolating mycelium. It seems to make sense that one wouldn't want many mycelia competing for the same substrate. Just curious if anyone knew for sure.


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A gram is better than a damn


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: HumboldtHort]
    #578254 - 03/14/02 12:05 AM (14 years, 8 months ago)

strange, I thought I said that in my last post. Must have forgotten:crazy:
Yes, professional growers use master cultures, as multispore spawn does not produce even flushes, which are essential in any commercial setup. Also some species will fruit very sparsely from a multi-strain casing. 


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hmmm........


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OfflineHumboldtHort
newbie
Registered: 02/13/02
Posts: 40
Last seen: 12 years, 8 months
Re: Agar Advantage? [Re: Champion des Champignons]
    #578261 - 03/14/02 12:15 AM (14 years, 8 months ago)

I was just reiterating. Cool. Thanks for the help. I appreciate it.


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A gram is better than a damn


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OfflineKindnug
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Male
Registered: 03/05/02
Posts: 123
Loc: Midwest, Go Bears
Last seen: 2 years, 4 months
Re: Agar Advantage? [Re: Champion des Champignons]
    #578990 - 03/14/02 07:50 PM (14 years, 8 months ago)

I was thinking of starting the same thing as well, doing some clones from the best fruiting cakes. I also wanted to try sectoring out and isolating a good single strain from print or whatever; hopefully just trying to judge by the types of myc growth which is best. Possible or no? I wanted to try and start this all up for around a hundred bucks, have you had any success with using homemade agar or have you been only been using store bought sterile agar? I was thinking of trying to make the agar myself with potatoes and the p/c. Everyone says its pretty easy to do, and for now I will only be doing p. cubensis anyway. I am not a big time grower or anything so this is purely for fun and to learn, I'm hoping that a while down the road it might pay off and give me even fruiting, consistent cakes to put in my pods. Right now it seems totally hampered by the multi-spore inocs, as far as cake surface area for cropping goes I am only getting flushes on maybe 60% of the surface, and that is the best cakes of the bunch. Typically I would say it's between 25-50%, it seems like the different looking myc growth sometimes doesn't produce anything. Being able to see the process of knotting to pinning to fruiting in the pod might have some advantage as far as seeing which substrain is best, any ideas how someone could take advantage of this? I was thinking maybe do some growth from a good fruiting clone in a petri, then sector out and colonize something (maybe double tall shot glasses or something full of coco-substrate? Grows very fast, 2-3 days for 100%) with each of the 10 or so substrains; because then you could hopefully study each seperately for traits, but the nice thing is they would share the same enviornment so that should not affect the cakes any differently. Then I'm hoping that whatever I find and eventually use to spawn more cakes will be already perfect for that enviornment, I wouldn't be judging petri dish growth versus cake growth, which could be far different I imagine depending on what it's growing on or whatever. Now I am a newbie so I may have something way off here. Any help is greatly appreciated, anything I ever get done that is different from what others have will certainly be shared. I'm going to start experimenting with different teks for printing soon too (once I get through 2 batches or so); so I will definitely hook up the FSR as soon as possible as well.... Who knows maybe someday I can be sending my own substrain of something to it for others to try!!! Peace out


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InvisibleChampion des Champignons
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Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: Kindnug]
    #579022 - 03/14/02 08:16 PM (14 years, 8 months ago)

I've just been using peroxidated wheatflour paste with good results.
I mix up some white flour with water until it is of a pourable consistency, pour into small jars (caviar jars are perfect:) to a depth of 3-4mm, pressure cook for 20 mins. It's important to let the p/c cool by itself because if you let the pressure off too quickly the "agar" bubbles up into a nice little loaf:)
Once cool I remove the lid, pour in enough 3% H2O2 to cover the surface, swirl it around for a few seconds and pour off the excess. The H2O2 is absorbed into the surface and creates a medium which seems pretty much impossible to contaminate, unless the peroxidated top layer is broken.
The peroxide means this is no good for culturing spores on, but if you have a flowhood/glovebox, you can miss out the peroxide. 


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hmmm........


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OfflineKindnug
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Registered: 03/05/02
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Re: Agar Advantage? [Re: Champion des Champignons]
    #579082 - 03/14/02 09:21 PM (14 years, 8 months ago)

So your techinique seems perfect for cloning in a normal everyday enviornment (exactly what I need). I think I will definitely try it first, seems far easier than homeade potato/yeast agar. And I already have everything too except something to put it in. Would this work if it was made up in qauntity in say a quart or pint jar, and then peroxidated as needed in the petris? This will not take any spores including a freshly made syringe right? Just a living tissue sample (clone or myc)? Is the idea to try and get the sample under the peroxidated layer or do you just throw it right on top and cover up? Do you conver tight to incubate or a little loose for air exchange? Thanks a lot for the info on your recipe, it doesn't get much easier than that!! Can't wait to try tomorrow if sterilization in a big jar will work. I imagine I'll probably have to pressure cook for 45 min, though. You think? Thanks again for all the help


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: Kindnug]
    #579131 - 03/14/02 10:39 PM (14 years, 8 months ago)

"Would this work if it was made up in qauntity in say a quart or pint jar, and then peroxidated as needed in the petris?"
No, it solidifies on cooking.
"This will not take any spores including a freshly made syringe right?"
Right.
"Is the idea to try and get the sample under the peroxidated layer or do you just throw it right on top and cover up?"
Just toss it on. Dipping the sample into h202 for a second seems to help it get used to the peroxidated medium faster than just dropping it on for some reason.
"Do you conver tight to incubate or a little loose for air exchange?"
I have the lids on tight to keep out any stray beasties, but seeing as how I open the jars every day or so for a peek air exchange is no prob.


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hmmm........


Edited by Champion des Champignons (03/14/02 10:41 PM)


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OfflineKindnug
member
Male
Registered: 03/05/02
Posts: 123
Loc: Midwest, Go Bears
Last seen: 2 years, 4 months
Re: Agar Advantage? [Re: Champion des Champignons]
    #580046 - 03/15/02 10:29 PM (14 years, 8 months ago)

Thanks for all the info man!!! You've been a great help, can't wait to give it a go this weekend.. Just picked up all the supplies, ready to try a few clones of these nice EQ's (great fruiting sustrain came out in 1 of my cakes) that are currently starting their second flush. Take it easy!!


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: Kindnug]
    #580128 - 03/16/02 12:22 AM (14 years, 8 months ago)

good luck. keep us updated...


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hmmm........


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OfflineKindnug
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Registered: 03/05/02
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Loc: Midwest, Go Bears
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Re: Agar Advantage? [Re: Champion des Champignons]
    #583899 - 03/20/02 04:29 AM (14 years, 8 months ago)

Sorry to post in an old thread, but I'm having trouble finding something to pressure cook my stuff in. I found these shallow jars online, I can't seem to get the picture to work but here is the explanation

Not a Door
You could pack pickled gherkins for lunch in these 3 oz. wide-mouth translucent white containers (polypropylene has good acid resistance) or you can use them in the lab. Foamed plastic liner in the opaque yellow plastic lid makes a tight seal. Overall, the jars are 1?1/2? high x 3?3/8? dia.
27150 POLYPROPYLENE JAR, 3 OZ $2.00 / PKG(6)

What do you think? Here is the link so you can see the pic if you want
http://www.sciplus.com/category.cfm?subsection=7&category=79

I couldn't get a direct link, you have to scroll down to where it says "more plastic bottles and jars" and click that, then do it again and it will be the first item. Titled, "Not a Door". I will be following your last post so I figured I'd check with you before ordering. I know that you won't be able to see inside the jar too well, but I plan to just use these for the first couple of sectors on 5 or 6 strains. I was thinking they are somewhat transluscuent, so when they've grown enough out I would only then open them. And as far as the p/c goes, I was thinking that I would build a aluminum rack to hold the jars further up in the cooker, away from the boiling water perhaps. And they will only be in for 20 min, anyway. You wouldn't think they would melt, would ya? Anyway after the first few sectors I would then transfer them over to petri with parafilm, once I had a decent idea of what I was working with. What do you think of this approach?


Edited by Kindnug (03/20/02 04:32 AM)


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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: Agar Advantage? [Re: Kindnug]
    #584255 - 03/20/02 03:10 PM (14 years, 8 months ago)

Those look perfect...if they resist autoclaving. Why not just buy a sleeve of disposable petris? They're pretty cheap and you don't _have_ to dispose of them.....:)


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hmmm........


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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