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OfflineF64
Sclerotia For Everyone!
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Registered: 06/06/11
Posts: 61
Last seen: 6 years, 1 month
Usefulness of agar use for growing sclerotia/philosophers stones?
    #16299268 - 05/29/12 02:07 AM (11 years, 8 months ago)

Hey Guys,
I'm all about growing sclerotia, and I now ordered prints of Mex A, with which I've had alright success previously when I ordered it in a spore syringe and created LC from that which I used to inoculate my jars.

Now I live somewhere else, and shipping a spore syringe is not an option. I don't have experience with either agar, nor spore prints.

Can agar help me..

1) get the spores inoculated (i.e. is doing it via agar better/safer than making a spore syringe straight from the print, and using the syringe for inoculation or an LC)

2) grow sclerotia faster/with more potency?

I'm especially interested in 2) - I know agar has considerable impact on cubes, I imagine mainly because they all grow at the same speed thus increasing harvest. But I think that that's not an issue with sclerotia, as there is not one set time for harvesting.

How can I use agar to select DNA that will grow faster/bigger/stronger sclerotia? Would it also be about selecting the strainier, rather than the fluffier mycelium?

Thanks guys!!! Looking forward to your advice :smile:


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InvisibleinskiM
Cortinariologist
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Registered: 02/28/06
Posts: 5,752
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Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: F64]
    #16299365 - 05/29/12 02:53 AM (11 years, 8 months ago)

If you can be sure that your spore print was made in sterile conditions you could make a syringe then use it to inoculate appropriate sterilised grain for sclerotia production, once you have harvested your sclerotia you could use agar to isolate a nice culture via tissue culture(cloning), just choose your nicest sclerotia then under sterile conditions excise a small piece from the inside of the stone and place it on agar.


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OfflineRogerRabbitM
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Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: F64] * 1
    #16299746 - 05/29/12 07:04 AM (11 years, 8 months ago)

Agar is very important in getting a good sclerotia producing strain.  You might find the agar prep video clip helpful in getting started.  After that, use the strain isolation video clip to isolate strains.  Rather than isolating for rhizomorphic mycelium, instead look for sectors of Mex a with brown coloring.  These tend to produce more sclerotia.  After a transfer or two, you should see actual sclerotia forming on the agar.  Grab these stones and transfer to new dishes.  When you have a culture which produces several stones on agar, transfer it to grains, and then do grain to grain transfers from there.  A good stone producing strain will be ready in two to three months.
RR


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"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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InvisibleLuger0815
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Registered: 12/21/10
Posts: 1,677
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Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: RogerRabbit]
    #16300232 - 05/29/12 10:22 AM (11 years, 8 months ago)

You can also set up some Multi Strain jars.
Select a good stone (one of the first stones) from this jars to clone the strain by transferring some flesh of the stone to an agar plate.
After the myc. expanded on the plate, transfer a piece of the myc into a Liquid Culture Jar.
Now you have a master LC Jar.
Store it in the fridge.
Use 1-2 ml from this Master-LC to make another LC Jar/s.
You are ready to set up hundreds of good Sclerotia producing Jars.

Just a suggestion.


Edited by Luger0815 (05/29/12 10:24 AM)


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Invisiblea social revolt
retired cultivator


Registered: 09/07/11
Posts: 833
Loc: Brooklyn, NYC Flag
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: Luger0815]
    #16300283 - 05/29/12 10:36 AM (11 years, 8 months ago)

:takingnotes:


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OfflineF64
Sclerotia For Everyone!
Female


Registered: 06/06/11
Posts: 61
Last seen: 6 years, 1 month
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: a social revolt]
    #16304486 - 05/30/12 02:02 AM (11 years, 8 months ago)

so helpful! thank you guys!


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OfflineRogerRabbitM
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Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: Luger0815]
    #16304969 - 05/30/12 06:47 AM (11 years, 8 months ago)

Quote:

Luger0815 said:

Store it in the fridge.





One needs to be very careful storing warm weather species under refrigeration.  I learned the hard way years ago that P mexicana will usually not survive a normal refrigerator, which operates only a few degrees above freezing.  I wouldn't keep it below about 50F.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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InvisibleLuger0815
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Registered: 12/21/10
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Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: RogerRabbit]
    #16305281 - 05/30/12 09:33 AM (11 years, 8 months ago)

Quote:

RogerRabbit said:
Quote:

Luger0815 said:

Store it in the fridge.





One needs to be very careful storing warm weather species under refrigeration.  I learned the hard way years ago that P mexicana will usually not survive a normal refrigerator, which operates only a few degrees above freezing.  I wouldn't keep it below about 50F.
RR





Oh, good to know.
Thanks for the advice!!!


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OfflineOwsley
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Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 1 month
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: RogerRabbit]
    #16305405 - 05/30/12 10:12 AM (11 years, 8 months ago)

Quote:

RogerRabbit said:
  Rather than isolating for rhizomorphic mycelium, instead look for sectors of Mex a with brown coloring.  These tend to produce more sclerotia.  After a transfer or two, you should see actual sclerotia forming on the agar.  Grab these stones and transfer to new dishes.  When you have a culture which produces several stones on agar...
RR




What do you do if your not seeing any "brown coloring"on agar?

Also I have some grain jars going with Mex A which just finished colonizing and again am not seeing any change in coloring or evidence that they are producing stones, just all completely white. How long would you wait and can I just attempt to fruit if they fail to produce stones?

And are the genetics, say if I took a spore print, then limited to a non-stone producing type?


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OfflineF64
Sclerotia For Everyone!
Female


Registered: 06/06/11
Posts: 61
Last seen: 6 years, 1 month
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: Owsley]
    #16311445 - 05/31/12 12:58 PM (11 years, 8 months ago)

Owsley, welcome in the sclerotia growing family :smile:. I grew around 15 jars of Mex A in several batches before.

Give the jars time! If they just finished colonizing, they may take another 1-3 weeks until you just start to notice a concentration of mycelium. For harvesting, you'll have to wait 2 months from colonization. I don't think there is a non-stone producing type of Mex A, and types derived from spore prints will certainly produce stones. If you need more help, PM me, as this is kind of off-topic.

Oh and also off-topic: I need a Mex A sporeprint badly! I now live on a different continent, and receiving a syringe here is just not an option. I want to spread the love of sclerotia in this region and help other people grow it, but I need one single print to get started. You don't happen to have one?


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InvisibleBobbySkidz
Aces High!


Registered: 02/10/12
Posts: 312
Loc: Neptune
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: RogerRabbit]
    #16315750 - 06/01/12 08:25 AM (11 years, 8 months ago)

Quote:

RogerRabbit said:
Quote:

Luger0815 said:

Store it in the fridge.





One needs to be very careful storing warm weather species under refrigeration.  I learned the hard way years ago that P mexicana will usually not survive a normal refrigerator, which operates only a few degrees above freezing.  I wouldn't keep it below about 50F.
RR




What would you recommend then for long term storage? Will it still go into stasis at 50, or keep growing?


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InvisibleLuger0815
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Registered: 12/21/10
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Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: BobbySkidz]
    #16322093 - 06/02/12 03:03 PM (11 years, 8 months ago)

Quote:

BobbySkidz said:
Quote:

RogerRabbit said:
Quote:

Luger0815 said:

Store it in the fridge.





One needs to be very careful storing warm weather species under refrigeration.  I learned the hard way years ago that P mexicana will usually not survive a normal refrigerator, which operates only a few degrees above freezing.  I wouldn't keep it below about 50F.
RR




What would you recommend then for long term storage? Will it still go into stasis at 50, or keep growing?




I store them at about 45F or 7C°, it seems that the myc survive it and doesn't keep on growing.......so far.:shrug:


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Offlinecaesar
Seeking causal knowledge


Registered: 05/08/12
Posts: 169
Loc: Texas
Last seen: 9 years, 1 month
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: Luger0815]
    #16325756 - 06/03/12 10:16 AM (11 years, 8 months ago)

Quote:

Luger0815 said:
Quote:

BobbySkidz said:
Quote:

RogerRabbit said:
Quote:

Luger0815 said:

Store it in the fridge.





One needs to be very careful storing warm weather species under refrigeration.  I learned the hard way years ago that P mexicana will usually not survive a normal refrigerator, which operates only a few degrees above freezing.  I wouldn't keep it below about 50F.
RR




What would you recommend then for long term storage? Will it still go into stasis at 50, or keep growing?




I store them at about 45F or 7C°, it seems that the myc survive it and doesn't keep on growing.......so far.:shrug:




How long have you kept a master culture? What is the best method/temperature for preserving master cultures for warm weather species?


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OfflineF64
Sclerotia For Everyone!
Female


Registered: 06/06/11
Posts: 61
Last seen: 6 years, 1 month
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: caesar]
    #16369812 - 06/12/12 09:44 AM (11 years, 8 months ago)

Quote:

I store them at about 45F or 7C°, it seems that the myc survive it and doesn't keep on growing.......so far.





Where I live, power is off 20% of the time, and sometimes days in a row. What would be the smartest way to keep a culture in normal room temperature? Grain jars, agar, LC?

Would a good beginner solution for selecting good genes be to select an early/large stone, clean it, and then just start a new LC or grain jar from it? The main drawback would be that myc with other genes would be transferred, and its probably slower than agar, right?


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Invisibledeucedbi9
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Registered: 10/24/06
Posts: 4,609
Loc: UK
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: F64]
    #16370216 - 06/12/12 11:05 AM (11 years, 8 months ago)

Quote:

F64 said:
Quote:

I store them at about 45F or 7C°, it seems that the myc survive it and doesn't keep on growing.......so far.





Where I live, power is off 20% of the time, and sometimes days in a row. What would be the smartest way to keep a culture in normal room temperature? Grain jars, agar, LC?






You could try drying spawn for later use. Reanimate on agar like this> Link

Obviously you would be more sterile about it to avoid the mess shown there.
You could just tip a small amount of spawn into a sterilised jar, in a glove box, before harvesting stones or spawning to bulk. The spawn will slowly dry out, through GE, and can be stored at room temperature for at least two years.


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Something tells me I'd better activate my prayer capsule


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InvisibleBobbySkidz
Aces High!


Registered: 02/10/12
Posts: 312
Loc: Neptune
Re: Usefulness of agar use for growing sclerotia/philosophers stones? [Re: deucedbi9]
    #16371889 - 06/12/12 04:35 PM (11 years, 8 months ago)

Damn good to know. Thanks...


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