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oakley
Stranger

Registered: 10/31/16
Posts: 55
Last seen: 2 years, 4 months
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Re: Cloning a Stone [Re: LocN9ne]
#23792659 - 11/01/16 07:28 PM (7 years, 2 months ago) |
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Quote:
AgarStudent said: With strain isolation, it looks like Chico is outperforming Galindoi 
Could you please elaborate on how you isolated that strain? What agar did you use for example? How many plates did you inoculate with spores? How many sectors did you isolate for a total number of petris? Did you use standard petris or ones with three sectors?
Quote:
LocN9ne said: Your ability to dig up specific relics on demand, and name drop... Combined with your post count and reg date really got me wondering who is pulling your strings there oakley
lel. thanks for the compliment.
Edited by oakley (11/01/16 07:45 PM)
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AgarStudent
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Registered: 01/25/15
Posts: 550
Last seen: 2 years, 10 days
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Re: Cloning a Stone [Re: oakley]
#23792741 - 11/01/16 07:46 PM (7 years, 2 months ago) |
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Absolutely -
1) Start with multispore. In this case I made up a syringe from the print of Chico Nindo.
2) Make up about 20 plates of Malt Extract Agar (MEA) on sterile 90mm plates over the flow hood.
3) Put a few drops around the centre of one, give it a few weeks to grow out. Take a look with a torch to look for denser growth or flecks of colour, rather than the fastest growing fuzzy mycelium (much like RR said).
4) Transfer the darker bits to another plate and let them grow out. Maybe do one more transfer and then drop a good size wedge in a jar or grains prepped with a bit of coffee.
--> At this stage I aren't 100% sure that my best jar is an isolate or a combination of sectors, but when the clone fills out I will keep you in the loop.
-------------------- Why reinvent the wheel when someone here has already done the science?
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oakley
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Registered: 10/31/16
Posts: 55
Last seen: 2 years, 4 months
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Quote:
AgarStudent said: 4) Transfer the darker bits to another plate and let them grow out. Maybe do one more transfer and then drop a good size wedge in a jar or grains prepped with a bit of coffee.
Honest thanks for the answer. So you did not wait for stones to form on agar is this correct? You just used the darker mycelium to inoculate your jars? I noticed that you're growing other sclerotia species too, where did you get your spores? Also, how did you build your flowhood?
Edited by oakley (11/01/16 08:06 PM)
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AgarStudent
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Registered: 01/25/15
Posts: 550
Last seen: 2 years, 10 days
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Re: Cloning a Stone [Re: oakley]
#23792846 - 11/01/16 08:14 PM (7 years, 2 months ago) |
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Please check our sponsors for spores.
I waited for stones to form on the agar, but only ever saw coloured bits and/or darker densities (light attenuating bits) on the plate.
The flowhood is just made from an inline exhaust fan (eBay), and a HEPA filter (eBay). The body you can build yourself from timber, but make sure it is well sealed etc.
I think I even found a better design when the one I got wears out.
http://www.freshcapmushrooms.com/learn/keeping-it-clean-how-to-design-and-build-a-laminar-flow-hood
-------------------- Why reinvent the wheel when someone here has already done the science?
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blackout


Registered: 07/16/00
Posts: 5,266
Last seen: 2 months, 25 days
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I have seen it recommended to look for stones & dark growth on agar, which makes sense, but I would not dismiss pure white growth.
A user called Aero cloned from a commercial stone. It was easily the fastest growing sclerotia I have ever seen in any thread, though I did not have the same results as him.
This was his thread https://www.shroomery.org/forums/showflat.php/Number/20833312/fpart/1/vc/1
In that thread you will see his growth appeared white, its hard enough to tell though as he grew in jars and most are backlit.
These are my clones of some of Aero's stones. I have posted the first 2 before but just found the third photo
this was number 1

and this was number 2,

This is the same number 2 4 days later.

I don't recall ever seeing any browning or hint of stones in these plates. I am starting it up again, from contaminated stones. My first was in a jar which was covered in agar inside so I do not have a clear photo. It was transfered to a glass petri and has no growth worth showing just yet.
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oakley
Stranger

Registered: 10/31/16
Posts: 55
Last seen: 2 years, 4 months
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Re: Cloning a Stone [Re: blackout]
#23799008 - 11/03/16 08:26 PM (7 years, 2 months ago) |
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Quote:
blackout said: A user called Aero cloned from a commercial stone.
I did not have the same results as him.
These are my clones of some of Aero's stones.
I don't recall ever seeing any browning or hint of stones in these plates.
It was transfered to a glass petri and has no growth worth showing just yet.
Maybe it's just senescence? I mean the commercial growers must have used G2G or some other method to create large amounts of myc to inoculate large amounts of grain fast, stones from that was cloned and grown on agar, and that was again grown on rye for many months, and stones harvested from that growth is what you're trying to isolate from. I think the myc is just too old at this point and white fluffy myc rather than colored myc might be a sign of it.
Blackout you were talking about introducing pieces of plastic into the growth medium for higher sclerotia production in the other thread, I searched through your posts and threads but couldn't find a post where you've said you tried your hypothesis, so I'm gonna ask here, did you ever get the chance to try that?
Also did anyone ever try to just add water into the jars as the substrate dried?
In the other thread
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Aero said: for stones im going to try to go bulk, and mix the grains with coir in a larger tray, one guy here did this and he said u just keep on watering the tray as it dries, and the sclerotia grows very fast and very big
he used these tupperware containers with the lid on, so there wont be enough evaporation to promote pinning but plenty of nutrition and water to get the sclerotia formation going
did anyone really try this out? I searched for it but couldn't find the thread aero was talking about. I'd appreciate if anyone could point me to said thread.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: Cloning a Stone [Re: oakley]
#23799028 - 11/03/16 08:31 PM (7 years, 2 months ago) |
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Not his specifically, but yeah I've seen TONS of sclerotia on pure CVG, done after picking nice fruits in lots of FAE. They look fuckin sexy, I'd totally do that
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stevo

Registered: 04/11/05
Posts: 5,100
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Re: Cloning a Stone *DELETED* [Re: Mad Season]
#23799802 - 11/04/16 06:12 AM (7 years, 2 months ago) |
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Post deleted by stevo
Reason for deletion: .
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oakley
Stranger

Registered: 10/31/16
Posts: 55
Last seen: 2 years, 4 months
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Re: Cloning a Stone [Re: stevo]
#23799940 - 11/04/16 07:44 AM (7 years, 2 months ago) |
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stevo said: I would clone one of those brown band like rhizomes connecting all of the stones together, and not try to expand it out and isolate to monoculture. I've cloned huge sclerotia before and got wimpy jar stones from the culture. So its not a guarantee. I think all the sclerotia strains are really the same species.
They are
http://www.mushroomjohn.org/Chapter-27-28.pdf
they can be considered different strains IMHO.
when you said you got small stones out of cloned stones, it reminded me of cyber's cloning efforts. perhaps the substrate they are grown on has a larger impact on some strains than others.
http://www.cyber-shrooms.com/growing/cloning-truffles-freshbox/
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blackout


Registered: 07/16/00
Posts: 5,266
Last seen: 2 months, 25 days
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Re: Cloning a Stone [Re: oakley]
#23803647 - 11/05/16 03:06 PM (7 years, 2 months ago) |
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oakley said: I think the myc is just too old at this point and white fluffy myc rather than colored myc might be a sign of it.
yeah this could be it, if its old it is still a very rapid producer of stones. I wonder if anyone who started with a "dark culture" ever took clones of clones of stones and found they were white.
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oakley said: Blackout you were talking about introducing pieces of plastic into the growth medium for higher sclerotia production in the other thread, I searched through your posts and threads but couldn't find a post where you've said you tried your hypothesis, so I'm gonna ask here, did you ever get the chance to try that?
I don't think I ever did, but I am pretty sure some other poster said they did. They either got the same growth or else a little more. If it was a lot more I definitely would have remembered that. I used to call them "interfaces" in threads if you are searching more and looking for a keyword.
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oakley said:Also did anyone ever try to just add water into the jars as the substrate dried?
Can't remember if I have done this, but brought it up a few times and some experienced growers thought it a good idea. I was later thinking coir in the mix might be a good idea as it will readily take on water again, while dried up grains may not. I also thought of having a layer of overhydrated rice at the bottom of a jar as a moisture source, it might not colonise but water could be absorbed from it.
I colonised white rice with Aeros culture and got only tiny stones after quite some weeks, I mixed this with coir to try and fruit them, I got no fruits at all but they did begin to produce stones. These were getting more moisture all the time with spraying and I will definitely be trying this again. The resultant stones contaminated easily so I would prefer to do it again under fully sterile conditions, injecting sterile water.
By weighing jars you can tell if any moisture has been lost, and have a better idea of how much to add.
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