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Offlinenimbus
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Sclerotia
    #2855757 - 07/04/04 09:04 AM (12 years, 5 months ago)

I ordered some fresh tampanensis sclerotia from holland, I want to use some tissue from it to go on growing my own. my questions are:

1- If I dry them with desiccant for long time storage, can I rehydrate a piece again (after 2 months) and put it on agar?

2- If I dont dry them, will they rot?

the problem is: I order it now and it will come fresh, but I can do the agar thing in two months. How can I succesfully preserve living tissue for 2 months?


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Offlinezeronio
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Re: Sclerotia [Re: nimbus]
    #2855769 - 07/04/04 09:17 AM (12 years, 5 months ago)

If you dry them then the chances that it will continue to grow on agar are small. I think that you should be able to keep them viable for 2 months if you put them into a sterile baggie and keep it in a refrigerator.


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Offlinenimbus
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Re: Sclerotia [Re: zeronio]
    #2855898 - 07/04/04 11:05 AM (12 years, 5 months ago)

i am planning to start some grass seed colonization this week (mexicana a), guess the tampanensis sclerotia will arrive by that time too. Can I just trow a piece of it (washed with H202 first?) i a sterilized jar/bag and start colonization?


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OfflinepsilocyberV
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Re: Sclerotia [Re: nimbus]
    #2855942 - 07/04/04 11:43 AM (12 years, 5 months ago)

You're likely to have some success with this method. I'd heard several reports of people being able to use harvested fresh sclerotia to inoculate sterile grain jars. You just need to make sure to give it a good H202 wash first (because chances are high that the exterior of the sclerotia is dirty as hell from handling). Work like you would when doing any other type of grain to grain transfer, in a sterile glove box or similar setting.


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Invisibletrade_om
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Re: Sclerotia [Re: zeronio]
    #2855992 - 07/04/04 12:16 PM (12 years, 5 months ago)

Quote:

zeronio said:
If you dry them then the chances that it will continue to grow on agar are small.



I disagree. Ive seen several species grown from re-hydratred sclerotia, that has been sitting dry in the fridge for more then 6 monthes.
2 important things to follow - dry with no heat and remember to rehydrate well before moving to agar. Even throw in sterilized water for 24hours prior to transfer to agar.
What I like to do is keep the sclerotia whole and intact. Dry it good with a decent dessicant. Then store in the fridge. When Im ready for transfering to agar I take my scalpel and slice the dried sclerotia in half. I then streak a small piece of the inner tissue of the sclerotia onto the scalpel and drop into some sterilized distilled water and let it sit for 24 hours. Then I pull it out with twizzers or scalpel, and drop onto agar plate (sometimes people also like to give a 2minute h2o2 dip but i dont always do it).
Hope this helps.


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Offlinenimbus
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Re: Sclerotia [Re: trade_om]
    #2856091 - 07/04/04 01:16 PM (12 years, 4 months ago)

thanks trade om, this was the way i was thinking would/could be right. Will try it too.
Anyone else other opinion?


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OfflineOpenminded
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Re: Sclerotia [Re: nimbus]
    #2856501 - 07/04/04 04:17 PM (12 years, 4 months ago)

Personally, what I would do is take a fresh sclerotium, cut it in half, scoop out a bit of tissue, dunk it in peroxide solution and then put it into karo water or similar liquid culture.
Then you can dry the rest of your sclerotia, and the mycelium growing in the liquid should be good for two months in the fridge (after letting it grow for a bit at room temp), and you can use that to innoculate grain.
From what I've heard/imagined, you'll have more luck innoculating grain with some mycelium from a liquid culture, rather than a lump of sclerotium, because the tissue in a lquid culture will be able to have a more intimate contact with your grain, and so should, I think, get growing better. Also it can be dispersed throughout the whole mass of grain, which should decrease colonisation time.
I'm getting some P. Mexicana A spores soon, and another thing I was thinking of trying was to use a little bit of sugar in the water that I mix with my rye grain, to give the spores/mycelium (depending on whether I go to grain directly from spores, or via a liquid culture) a bit of readily available food which it can use while it begins to attack the grain which, being solid, I imagine is a less readily accessable food supply.


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Offlinenimbus
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Re: Sclerotia [Re: Openminded]
    #2856666 - 07/04/04 05:29 PM (12 years, 4 months ago)

nice idea but I have only 1 week of time to be able to do something, next week I am leaving home for 8 weeks, so I want to start a colonization this week. for this reason the best choice is innoculating with a piece of sclerotia i think..

what do you think anybody?


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Invisiblematts
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Re: Sclerotia [Re: nimbus]
    #2856755 - 07/04/04 06:21 PM (12 years, 4 months ago)



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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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