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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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Petri dish cultures and sectors
    #8099268 -

I experimented with peroxide agar and am having a hell of a time with it. My problem/question is as follows. Mycelium is having a hard time colonizing the agar. Grant it, I have no contamination whatsoever it took forever for the mycelium to really take hold of the agar. I identified 5 sectors on that dish. I then transfered a tiny bit of each sector to 5 different dishes. Now those tiny individual slices are also producing 3 to 5 sectors! The problem here is that since the mycelium is growing at a snails pace, I can't say that I can see any outstanding features from any of those sectors! No agressive growth, no fantastic rhizomorhic growth. Just slow steady fuzzy growth. If I transfer the sectors to other dishes, I will have 15 dishes going with no real clue as to which one is the one I want to transfer to grain. Furthermore, if those 15 isolates produce 3 to 5 sectors I'm screwed! What would you guys do?


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AMU Q&A thread.

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Re: Petri dish cultures and sectors [Re: The shroomy 1]
    #8099906 -

Keep going until you get single sectors then fruit them all. Keep the cultures of the best performers.

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Re: Petri dish cultures and sectors *DELETED* [Re: The shroomy 1]
    #8100017 -

Post deleted by Morelman

Reason for deletion: Never again...



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Re: Petri dish cultures and sectors [Re: Morelman]
    #8100123 -

Agreed. Peroxidated agar kills growth rates. It usually takes three to four transfers to get single sectors. Isolate all of them to determine the best performers. By the third transfer, you should see some rhizomorphic mycelium.
RR


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Re: Petri dish cultures and sectors [Re: RogerRabbit]
    #8103936 -

Thanks guys! I still have 8 dishes of peroxide agar left. I'm going to decide on the 8 best looking fuzzy sectors and get those going. That will free up the rest of the petri dishes. I'll get some straight agar going to re-fill the petri's and maybe the next go around will reveal better traits. Once again, Thanks for the input!

The Shroomy 1


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Re: Petri dish cultures and sectors *DELETED* [Re: The shroomy 1]
    #8105121 -

Post deleted by Morelman

Reason for deletion: Never again...



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Re: Petri dish cultures and sectors [Re: Morelman]
    #8105958 -

MorelMan, I feel your pain in the area of throwing away cultures.  I recently had to dump over 20 agar plates of oysters in the trash to make room.  I mean, come on, there just oysters, right?  But even trying to tell myself that just simply kills. 

And if you're having a hard time throwing them away, I know of a good home for adoption...:tongue:

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Re: Petri dish cultures and sectors [Re: Morelman]
    #8108781 -

Morelman said:
By "straight agar" I assume you mean straight PDA or MEA or some other nutritious formula. Straight agar in the literal sense won't work for what you're trying to accomplish.


LOL! Of course I don't mean it in the literal sense! I just meant that I won't be putting peroxide in it. I'm thinking of purchasing media antibiotic from workman. I'm thinking it won't be so harsh on the mycelium.


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Re: Petri dish cultures and sectors *DELETED* [Re: The shroomy 1]
    #8108918 -

Post deleted by Morelman

Reason for deletion: Never again...


Edited by Morelman (03/05/08 10:59 PM)

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Re: Petri dish cultures and sectors [Re: Morelman]
    #8111139 -

How about heating your remaining peroxidated petris to destroy the peroxide? From memory 70 degrees Celsius will do it without melting your plates. I've tried it for getting spores going on spare H2O2 plates.


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Re: Petri dish cultures and sectors [Re: worowa]
    #8113644 -

Thanks for the advice guys! I'm going to do the transfers tomorrow and get some more agar plates prepped.


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Re: Petri dish cultures and sectors [Re: The shroomy 1]
    #8136530 -

Someone should have asked what type of mushroom you are culturing on the peroxidated plates. I would really like to know.

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Re: Petri dish cultures and sectors [Re: The shroomy 1]
    #8179055 -

From what i've read, sectoring is increased by availability of starch - i.e from exploded rye kernels. you could try adding a little to your recipie for the first plates if you want to isolate a rhizo sector quicker.

definately drop the h2o2 since its clearly slowing your culture down. Nothing worse than having things impeding an allready slow process! lol

good luck

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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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