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AgarStudent
Noob


Registered: 01/25/15
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Cloning a Stone
#23787576 - 10/31/16 05:53 AM (7 years, 2 months ago) |
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I have this one jar of Mexicana that has progressed really well, except I no longer have the agar plate that it came from.
I don't want to ruin the jar, but at the same time would like to take a sample of a stone and grow it out on agar to grow more jars the same? I figure whatever sectors that are present in the jar should likely be present in the stone?
Is is possible to clone a stone and not ruin the jar if I do it over the flow hood, use a flame sterilised scalpel and put the lid back on afterwards?
Thanks guys
-------------------- Why reinvent the wheel when someone here has already done the science?
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blindingleaf
blue collar underworld


Registered: 07/19/13
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yea, possible (albeit weird) but depending on where the stone is in the jar….difficult.
hopefully its close to the top
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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AgarStudent
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Yep there are stones everywhere:
-------------------- Why reinvent the wheel when someone here has already done the science?
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enlightenment
alchemist


Registered: 08/09/09
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Nice jar. Packet with sclerotia. You can wait until you harvest all and clone a stone at this moment.
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blindingleaf
blue collar underworld


Registered: 07/19/13
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go for it 
TBH, u could just use a new scalpel blade and kinda "slice" a piece of one of the stones on the glass. its sterile, so u don't need to worry about the outside of the "fruit body" being contaminated
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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AgarStudent
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Okay, thanks guys. Will give it a go
-------------------- Why reinvent the wheel when someone here has already done the science?
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LocN9ne
ɢᄋᄋd ԲᄋЯ ᄁᄋȚᅢΙᄁɢ ᄂᄋ₩ᄂΙԲᄐ



Registered: 04/17/15
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Loc: to the brain
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You dont even have to dig a stone out... Stab a piece of grain and throw that bitch onto a plate... But either way is fine.
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Q&A US vs. THEM The more I learn, the less I know.
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sandy_vag
Keep Dreaming



Registered: 06/14/16
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Re: Cloning a Stone [Re: LocN9ne]
#23787978 - 10/31/16 09:39 AM (7 years, 2 months ago) |
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Anybody seen mexicana A-strain spores around? Can't locate them at any USA vendors.
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LocN9ne
ɢᄋᄋd ԲᄋЯ ᄁᄋȚᅢΙᄁɢ ᄂᄋ₩ᄂΙԲᄐ



Registered: 04/17/15
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Atl7? It'll be called galindoi
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Q&A US vs. THEM The more I learn, the less I know.
Edited by LocN9ne (10/31/16 11:10 AM)
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oakley
Stranger

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Re: Cloning a Stone [Re: LocN9ne]
#23788257 - 10/31/16 11:19 AM (7 years, 2 months ago) |
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If you used an isolate from the agar plate you can just drop any grain from your jar on agar since all the mycelium in the jar would have grown from a single genetic lineage.
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LocN9ne
ɢᄋᄋd ԲᄋЯ ᄁᄋȚᅢΙᄁɢ ᄂᄋ₩ᄂΙԲᄐ



Registered: 04/17/15
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Re: Cloning a Stone [Re: oakley]
#23788319 - 10/31/16 11:44 AM (7 years, 2 months ago) |
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True, and on the other end of the spectrum if it was from MS the diversity would be so great that you will get pretty much around the same results with the same single grain method you just mentioned... Isolates have a place, but they are over rated by folks until they learn quite a bit more... Isolates arent anywhere near being in my arsenal yet... Too much other shit to nail down first,
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Q&A US vs. THEM The more I learn, the less I know.
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Spore Ninja
PsychoMycoPhile


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Quote:
sandy_vag said: Anybody seen mexicana A-strain spores around? Can't locate them at any USA vendors.
I believe Sporeworks was the originator of the Mexi A line, so I'd go ahead and invest in the good genetics they have.
Plan B would be to order some truffle from one of our vendors that offers them, and then clone them. That's what I'm doing to get some good Tamapanenis genetics. It's about $20 if you don't use the registered/tracked shipping option- $40 if you do.
-------------------- I'm interested in Dr. Pollock's work and his (especially older) strains. Anyone have pix of his Shroommobile RV?? PSA: Protonmail offers free, encrypted email hosted in privacy loving Switzerland. Red Phone / Signal offer free encrypted phone calls and texting between users. All of these programs are available free for PC and Android or apple and are available in the respective stores or online by googling the names. Stay safe out there! The Electronic Freedom Foundation's review of text and messaging program's security is here>>> https://www.eff.org/node/82654
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blackout


Registered: 07/16/00
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Quote:
Spore Ninja said:
Quote:
sandy_vag said: Anybody seen mexicana A-strain spores around? Can't locate them at any USA vendors.
I believe Sporeworks was the originator of the Mexi A line, so I'd go ahead and invest in the good genetics they have.
+1
Quote:
Spore Ninja said: Plan B would be to order some truffle from one of our vendors that offers them, and then clone them.
I presume he is in the US if looking for US vendors. I would not be ordering truffles to the US, I thought no shroomery sponsors would send truffles to the US either. I meant to mention it in your other thread.
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sandy_vag
Keep Dreaming



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Re: Cloning a Stone [Re: blackout]
#23788799 - 10/31/16 02:34 PM (7 years, 2 months ago) |
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Yea I been checking sporeworks for over a year now and they've always been out of stock on the mexicana a-strain. Even sent an email to ask if they would get any more but never heard back.
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AgarStudent
Noob


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Ah yeah I have no idea if it is an isolate in that jar or not. I think RR mentioned that he would try to transfer the brown bits for better stones:
Quote:
RogerRabbit said: The darker parts produce more stones. This is just an observation. Isolate some of both to confirm. RR
I guess if it grows out symmetrically and does not have sectors then it may be an isolate. Still different sectors may work together to produce stones too.
I am confused a bit by the naming convention, but to get things straight:
Psilocybe Mexicana: Strain A = 'Chico Nindo' and;
Psilocybe Mexicana: Strain B = 'Jalisco'
Psilocybe Galindoi: = ATL#7
Could anyone clarify that this is right or if there are any bits I have missed? 
One other thing.. What do you do with jars like this that are about three months old, with different genetics and still look kinda crappy? Do you try and fruit them or leave them?

Cheers
-------------------- Why reinvent the wheel when someone here has already done the science?
Edited by AgarStudent (10/31/16 06:44 PM)
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LocN9ne
ɢᄋᄋd ԲᄋЯ ᄁᄋȚᅢΙᄁɢ ᄂᄋ₩ᄂΙԲᄐ



Registered: 04/17/15
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Loc: to the brain
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That jar is fucked with bacteria, i dont see a single stone in there... You could attempt to spawn it, but three months of bacteria breeding is not a promising prospect.
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Q&A US vs. THEM The more I learn, the less I know.
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AgarStudent
Noob


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Re: Cloning a Stone [Re: LocN9ne]
#23789665 - 10/31/16 07:29 PM (7 years, 2 months ago) |
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Alright, that sounds possible. Maybe the brown bits are really tiny stones trying to co-exist with the bacteria and somehow they have achieved a balance? Still I guess it is not useful for anything but the compost bin? 
Edited by AgarStudent (11/01/16 06:03 AM)
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oakley
Stranger

Registered: 10/31/16
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Last seen: 2 years, 4 months
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Quote:
AgarStudent said: I am confused a bit by the naming convention, but to get things straight:
Psilocybe Mexicana: Strain A = 'Chico Nindo' and;
Psilocybe Mexicana: Strain B = 'Jalisco'
Psilocybe Galindoi: = ATL#7
Could anyone clarify that this is right or if there are any bits I have missed?
This is absolutely not correct. Mex-A and Mex-B have been isolated by workman I think, and jalisco have been domesticated by workman
https://www.shroomery.org/forums/showflat.php/Number/4153712
chicon nindo was collected by azure and distributed by azure/elfstone. the thread is here on shroomery.
https://www.shroomery.org/forums/showflat.php/Number/15954278
additionally there is also a lox strain going around collected by alan rockerfeller and distributed by prof pinhead.
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LocN9ne
ɢᄋᄋd ԲᄋЯ ᄁᄋȚᅢΙᄁɢ ᄂᄋ₩ᄂΙԲᄐ



Registered: 04/17/15
Posts: 7,076
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Re: Cloning a Stone [Re: oakley]
#23792427 - 11/01/16 06:32 PM (7 years, 2 months ago) |
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Quote:
oakley said:
Quote:
AgarStudent said: I am confused a bit by the naming convention, but to get things straight:
Psilocybe Mexicana: Strain A = 'Chico Nindo' and;
Psilocybe Mexicana: Strain B = 'Jalisco'
Psilocybe Galindoi: = ATL#7
Could anyone clarify that this is right or if there are any bits I have missed?
This is absolutely not correct. Mex-A and Mex-B have been isolated by workman I think, and jalisco have been domesticated by workman
https://www.shroomery.org/forums/showflat.php/Number/4153712
chicon nindo was collected by azure and distributed by azure/elfstone. the thread is here on shroomery.
https://www.shroomery.org/forums/showflat.php/Number/15954278
additionally there is also a lox strain going around collected by alan rockerfeller and distributed by prof pinhead.
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
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Q&A US vs. THEM The more I learn, the less I know.
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AgarStudent
Noob


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Re: Cloning a Stone [Re: LocN9ne]
#23792447 - 11/01/16 06:38 PM (7 years, 2 months ago) |
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With strain isolation, it looks like Chico is outperforming Galindoi
-------------------- Why reinvent the wheel when someone here has already done the science?
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oakley
Stranger

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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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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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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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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
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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
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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
Quote:
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

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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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Quote:
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


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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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Quote:
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.
Quote:
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.
Quote:
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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