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InvisibleZwieback0
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Isolating on Agar
    #2425294 - 03/12/04 08:02 PM (20 years, 20 days ago)

After letting the mycelium take over the agar, how do you determine which chunk of agar to isolate?

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Invisiblebathrobe
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Re: Isolating on Agar [Re: Zwieback0]
    #2425325 - 03/12/04 08:13 PM (20 years, 20 days ago)

don't let it take over all the way. just let it colonize about 3/4 and take the fastest growing stuff. keep records and keep the old agar plates in the fridge so that you know which strains fruit the best, as well.


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OfflinePsilygirl
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Re: Isolating on Agar [Re: Zwieback0]
    #2425374 - 03/12/04 08:31 PM (20 years, 20 days ago)

just take a wedge shaped portion of the strongest, fastest healthiest looking mycelium.


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Anonymous

Re: Isolating on Agar [Re: bathrobe]
    #2425415 - 03/12/04 08:44 PM (20 years, 20 days ago)

See as soon as you see lil fluff balls seperate to their own plate.
Then look for ropey nonarial strands( see pic of annos GT) (fluff on one side rhizomorphic ropey strands on the other) of mycelium that is running fast outwerds away from the cottony fluff ball. When you have them sectioned they should grow in a perfect circle evenly from center out. If you dont see that you have more then one substrain on your plate. When you see ropey mycelium that means you have a mated pair of monokaryons.Which results in what is called a dikaryon which means mycelium capable of fruiting.

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InvisibleZwieback0
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Re: Isolating on Agar [Re: ]
    #2425466 - 03/12/04 09:12 PM (20 years, 20 days ago)

So I'm looking for ropey, correct? Thanks

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Anonymous

Re: Isolating on Agar [Re: Zwieback0]
    #2426662 - 03/13/04 07:54 AM (20 years, 20 days ago)

yes.

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InvisibleZwieback0
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Re: Isolating on Agar [Re: ]
    #2427221 - 03/13/04 11:36 AM (20 years, 20 days ago)

Seems easy enough! :smile:

What way should I use to preserve the final agar?  I've heard that you can just take a chunk and throw it in a vial of sterilized water and cap it.

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Anonymous

Re: Isolating on Agar [Re: Zwieback0]
    #2427353 - 03/13/04 12:19 PM (20 years, 20 days ago)

Colonized agar can be kept indefinitely when coated with sterile mineral oil and refrigerated at 37 - 41F as per Stamets' TMC...

Edited by Anonymous (03/13/04 12:20 PM)

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Offlinedebianlinux
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Re: Isolating on Agar [Re: Zwieback0]
    #2434807 - 03/15/04 12:06 PM (20 years, 18 days ago)

Quote:

Zwieback0 said:
So I'm looking for ropey, correct? Thanks




I'd like to point out that I have seen numerous references both by Stamets and some members here (Joshua for sure) that while ropey (aka rhizomorphic) is a very good indicator it is not the be all end all. In fact, cottony (aka tomentose) is not, by necessity, an indicator of weaker pairs/growth/etc. I have seen several examples of tomentose culture proving to be very vivacious. I'm certain Joshua created a thread concerning just that subject.

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Anonymous

Re: Isolating on Agar [Re: debianlinux]
    #2434822 - 03/15/04 12:09 PM (20 years, 18 days ago)

Thats is definetly case for azuresence and cyanesence.

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InvisibleMrMaddHatter
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Re: Isolating on Agar [Re: ]
    #2435158 - 03/15/04 01:42 PM (20 years, 18 days ago)

Agar work is so much fun. :loveeyes:




God help me. I've become a freakin nerd.

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InvisibleJoshua
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Re: Isolating on Agar [Re: debianlinux]
    #2435375 - 03/15/04 02:49 PM (20 years, 18 days ago)

I have seen isolates perform with varying degrees in different agar plates, all the same recipe or even the same batch.

I think often there are growth conditions that lead to a prevailence of rhizomorphic growth.

I do trust what Stamets has to say as his experiences with agar cultures make mine look miniscule.

Joshua


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Invisibletripndicular
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Re: Isolating on Agar [Re: MrMaddHatter]
    #2435427 - 03/15/04 03:00 PM (20 years, 18 days ago)

Quote:

MrMaddHatter said:
Agar work is so much fun. :loveeyes:




God help me. I've become a freakin nerd.




I agree agar is very fun , and is not for only the experienced , never let anyone tell you that .
The idea of getting a good issolate is to :
Harvest the fastest , strongest looking myc that is not sectoring , in most cases rhizomorphic stranding is what one looks for . It is not always necessarily the best , key words are fastest , strongest , non sectoring myc . Species and type of agar used can and most times determines what type of and health of myc one sees growing in the petri . No never wait till myc reaches edges of dish to select an issolate , it should be done just after 3\4 of the dish are dominated by myc .
Here are a couple of pics of some petris after only one issolation was chossen .



Good Luck


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OfflineRogerRabbitM
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Re: Isolating on Agar [Re: Zwieback0]
    #2438548 - 03/16/04 08:43 AM (20 years, 17 days ago)

Here's a link to a strain isolation tek I did over at Mycotopia.
http://www.mycotopia.net/cgi-bin/discus/show.cgi?tpc=5&post=133501#POST133501


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