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Dip41ea
Stranger

Registered: 12/22/13
Posts: 50
Last seen: 6 years, 1 month
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Selecting sclerotia isolate on agar - WITH PICS
#21347993 - 03/01/15 12:16 PM (9 years, 2 months ago) |
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I am new to sclerotia forming mushrooms. Using a spore print, I knocked up some agar plates. They have all germinated with what appears to be healthy growth. NOW, how do I select which portions to isolate and transfer to another dish, or is it ok to take the best plate and just go straight to some RB jars?
My goal is sclerotia, not fruiting. I am under the impression (i.e., read in other threads) that it is best to select brownish sectors of the agar or where it appears that stones my be forming. None of the plates present as stark/clear stones on agar as I have seen in prior threads. The plates are now 3 weeks--the slow growth is assumedly due to cold temperature (high 60s-70), which is normal for me.
Looking at the below pictures, which plates do you recommend for transfer (to new plates or RB jars)? I have circled the portions of the dishes that, to my novice eye, looking promising.
   
The pics are best if enlarged. I'm leaning towards throwing plates 2 and 3 into different RB jars and seeing what happens and pitching the rest of the plates. Or should I transfer to a new plate first (or select some other plates)?
Please help! All suggestions appreciated!!
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urthtown
meat popsicle



Registered: 10/26/13
Posts: 1,039
Loc: Eastern Canadia
Last seen: 2 years, 7 months
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Re: Selecting sclerotia isolate on agar - WITH PICS [Re: Dip41ea]
#21348020 - 03/01/15 12:22 PM (9 years, 2 months ago) |
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You could transfer a wedge to a small container of grain or whatever nutritious sub you want really and wait for the first sclerotia to form then harvest and clone that guy. I'm doing exactly this right now from a P. galindoi MSS:

Clone a stone
-------------------- Cluster Headache sufferer? Cluster Busting Veil Tear GIF Flower Pot Grow GIF Mini Mono Tub GIFS "All mushrooms are edible, but some only once." -- Croatian Proverb
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Dip41ea
Stranger

Registered: 12/22/13
Posts: 50
Last seen: 6 years, 1 month
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Re: Selecting sclerotia isolate on agar - WITH PICS [Re: urthtown]
#21348087 - 03/01/15 12:40 PM (9 years, 2 months ago) |
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Thanks. Is selecting a brownish sector / stone on agar just about trying to get *more* stones in the jar? I.e., do I run a risk of no stones forming when I go to grain if I don't find a brown'ish agar edge to transfer? Also, from which plate do you think I should get the wedge?
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filthyknees
no coincidence


Registered: 03/08/13
Posts: 6,283
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Re: Selecting sclerotia isolate on agar - WITH PICS [Re: Dip41ea]
#21348320 - 03/01/15 01:44 PM (9 years, 2 months ago) |
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I agree that 2 and 3 look like good candidates. When I transfer stones they pop off the agar, no cutting needs to be done ime. Three o'clock on 3 looks like a stone about to form. Hope you'll report back
-------------------- But if you're in a hurry, and really got to go If you're in a hurry, might have to find out slow That it's one thing to try and another to fly You get there quicker just a step at a time It's one thing to bark, another to bite The show ain't over till you pack up at night
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urthtown
meat popsicle



Registered: 10/26/13
Posts: 1,039
Loc: Eastern Canadia
Last seen: 2 years, 7 months
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Re: Selecting sclerotia isolate on agar - WITH PICS [Re: Dip41ea]
#21348341 - 03/01/15 01:49 PM (9 years, 2 months ago) |
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I would just grab a healthy wedge from that third plate and nocc up a half pint of WBR. Then when a stone formed after 100% colonization I would pop the cake out, clone the stone and then case the cake back into its container to see if I could get some spore prints from it. Then you have more prints to have and trade and a clone of a solid stone producer. It won't be an isolate right away but you could proceed to make transfers and try to isolate it after that. At least that way you would be starting with known stone producing strains.
But I'm no expert, maybe Stonesun will comment
-------------------- Cluster Headache sufferer? Cluster Busting Veil Tear GIF Flower Pot Grow GIF Mini Mono Tub GIFS "All mushrooms are edible, but some only once." -- Croatian Proverb
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Abshroom
Cause he can


Registered: 06/05/14
Posts: 810
Loc: Germany
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Re: Selecting sclerotia isolate on agar - WITH PICS [Re: urthtown]
#21348562 - 03/01/15 02:51 PM (9 years, 2 months ago) |
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Quote:
urthtown said: You could transfer a wedge to a small container of grain or whatever nutritious sub you want really and wait for the first sclerotia to form then harvest and clone that guy. I'm doing exactly this right now from a P. galindoi MSS:

Clone a stone 
 Tomorrow I will crumble up some mexicana A grain jars and harvest my first tiny stones. Does it matter how far the stone has already grown? IMO the younger the stone is, the better? So a harvest when the first stones are forming would be better then waiting 6 months and clone a giant full matured stone?
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urthtown
meat popsicle



Registered: 10/26/13
Posts: 1,039
Loc: Eastern Canadia
Last seen: 2 years, 7 months
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Re: Selecting sclerotia isolate on agar - WITH PICS [Re: Abshroom]
#21348584 - 03/01/15 02:57 PM (9 years, 2 months ago) |
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Uh I don't think it really matters but I would harvest and clone a stone after only a month if only for expedience but I'm sure you could successfully clone a 3 or even 6 month old stone in its first generation of growth from a MS.
-------------------- Cluster Headache sufferer? Cluster Busting Veil Tear GIF Flower Pot Grow GIF Mini Mono Tub GIFS "All mushrooms are edible, but some only once." -- Croatian Proverb
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Fungus Mountain
Poke-N-Squirt



Registered: 10/16/14
Posts: 1,188
Loc: Front row @ the Freakshow
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Re: Selecting sclerotia isolate on agar - WITH PICS [Re: Dip41ea]
#21348690 - 03/01/15 03:27 PM (9 years, 2 months ago) |
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You won't know if you have sclerotia forming genetics on agar, until you actually get stone formation and transfer to another dish > grains.
You are light-years away from an isolate, much less one that you know will be a stone producer. The only way to achieve that will be more transfers. Short of doing it that way, do as Urthtown suggests and try and knock up a cake or small grain jar and hope that you grow a stone or 2, then clone it.
-------------------- “Until they became conscious they will never rebel, and until after they have rebelled they cannot become conscious.” ― George Orwell, 1984 "If you can't explain it simply, you don't understand it well enough." ― Albert Einstein
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