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yisroel
Stranger
Registered: 09/27/14
Posts: 4
Last seen: 10 years, 2 months
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Trying cultivation from spores, need input. (photos inside)
#20625019 - 09/27/14 09:58 AM (10 years, 3 months ago) |
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Used a paper napkin to wipe off some spores from a dish. Liquefied that and strained to get a 'spore syringe'. In a 'clean' room tried to inoculate some jars. Here are the results after 1 week incubation at 84 deg.
Jars with rye seed prepared and pc'ed for 60 minutes. Used some foam on the jar lids.



Next photos are from beer agar petri dishes. Boiled a beer, added agar, pc for 60 min. Inoculated with 'white tissue' from the area of the 'root' hidden in dirt, from a live shroom specimen.


I am practicing my 'proper clean room' techniques, while I wait to get a decent spore print or spore syringe.
I have read that even if you inoculate a petri dish with contaminated spore syringe, and bacteria growth occurs it is possible to isolate a single 'hair' of mycelium and transport it to a clean petri dish for cultivation. How do I tell mycelim from other 'whitish' growth? Still learning.
Thank you for your advice and patience.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625101 - 09/27/14 10:20 AM (10 years, 3 months ago) |
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all that shit is totally fucked throw it away.
Quote:
Used a paper napkin to wipe off some spores from a dish. Liquefied that and strained to get a 'spore syringe'. In a 'clean' room tried to inoculate some jars. Here are the results after 1 week incubation at 84 deg.
Jars with rye seed prepared and pc'ed for 60 minutes. Used some foam on the jar lids.
napkins are not sterile, you need to work with sterile tools when working with spores. Inoculation loops get flamed until red hot to sterilize before ever touching spores.
there's no clean room. you need a perfectly sterile environment like an ISO 1 clean room even in an ISO 2 clean room you would need to work in a still air box or in front of a flow hood. any amount of cleaning is completely futile you have to work sterile with sterile procedures not in a "clean" room. spraying a room down with lysol or alcohol or bleach is the biggest noob mistake there is.
84F is way too hot, incubation should occur at room temperature and never above 75F. from 75-81F cubensis mycelium moves as fast as it's genes are going to let it move. After 81F the mycelium will actually stall or slow and at temps above 75F you're drastically increasing the ability of contamination. 68-72F is ideal.
you have to PC rye for at least 90M at 15psi 60m won't cut it.
that foam on your jar lid is good for making a yeast slurry to pitch into beer but for mycology work it's not going to cut it either.
Edited by Trusted cuItivator (09/27/14 10:25 AM)
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yisroel
Stranger
Registered: 09/27/14
Posts: 4
Last seen: 10 years, 2 months
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Re: Trying cultivation from spores, need input. (photos inside) [Re: bodhisatta]
#20625111 - 09/27/14 10:23 AM (10 years, 3 months ago) |
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Quote:
bodhisatta said: all that shit is totally fucked throw it away.
Lel. Throwing is easy. Could you please be more specific?
What can you tell me about the petri dish?
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pawnzy
That Guy


Registered: 03/04/14
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Last seen: 7 years, 9 months
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625128 - 09/27/14 10:29 AM (10 years, 3 months ago) |
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Quote:
yisroel said:
Quote:
bodhisatta said: all that shit is totally fucked throw it away.
Lel. Throwing is easy. Could you please be more specific?
What can you tell me about the petri dish?
It is easy, no need for specifics. What he means by fucked is its A) not going to work B) Infested with contamination C) fucked.
I dont know where you got that paper towel idea but  ? The petri dish: why did you boil a beer? Why not use a proper and recommended agar recipe
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Trying cultivation from spores, need input. (photos inside) [Re: pawnzy]
#20625133 - 09/27/14 10:30 AM (10 years, 3 months ago) |
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there's nothing to save from that petri dish it's covered in bacteria there's no possible way to isolate anything clean from that it's good as trash.
beer has isohumulone it's a mold inhibitor and mushrooms are mold. granted cubensis can likely grow on most beers it's still a bad idea.
brewers yeast are a genetically modified organism over centuries and that's why they can propagate in a beer full of iso-acids
Edited by Trusted cuItivator (09/27/14 10:32 AM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625136 - 09/27/14 10:31 AM (10 years, 3 months ago) |
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The petri looks like a contam buffet, lots of bacteria and some mold too. Might be some cube myc but good luck getting it clean. It could be done but is not likely worth the effort. Start researching proper sterile tek and throw all that stuff away.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Trying cultivation from spores, need input. (photos inside) [Re: Pastywhyte]
#20625157 - 09/27/14 10:37 AM (10 years, 3 months ago) |
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Before you try again do a lot of research. A lot of research. A lot of research.
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Stromrider
This must be the place



Registered: 06/02/13
Posts: 7,350
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Re: Trying cultivation from spores, need input. (photos inside) [Re: Pastywhyte]
#20625161 - 09/27/14 10:38 AM (10 years, 3 months ago) |
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Trying cultivation from spores, need input. (photos inside) [Re: Stromrider]
#20625166 - 09/27/14 10:40 AM (10 years, 3 months ago) |
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not to be a dick but if I were to post a how not to do things, I would have done exactly this to the T.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625169 - 09/27/14 10:41 AM (10 years, 3 months ago) |
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Quote:
yisroel said: Used a paper napkin to wipe off some spores from a dish. Liquefied that and strained to get a 'spore syringe'. In a 'clean' room tried to inoculate some jars. Here are the results after 1 week incubation at 84 deg.
Jars with rye seed prepared and pc'ed for 60 minutes. Used some foam on the jar lids.



Next photos are from beer agar petri dishes. Boiled a beer, added agar, pc for 60 min. Inoculated with 'white tissue' from the area of the 'root' hidden in dirt, from a live shroom specimen.


I am practicing my 'proper clean room' techniques, while I wait to get a decent spore print or spore syringe.
I have read that even if you inoculate a petri dish with contaminated spore syringe, and bacteria growth occurs it is possible to isolate a single 'hair' of mycelium and transport it to a clean petri dish for cultivation. How do I tell mycelim from other 'whitish' growth? Still learning.
Thank you for your advice and patience.
just in case OP decides to delete his post as often happens.
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yisroel
Stranger
Registered: 09/27/14
Posts: 4
Last seen: 10 years, 2 months
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625205 - 09/27/14 10:50 AM (10 years, 3 months ago) |
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Research by itsef in not sticking in my mind, I need to practice. This is it. So far I havent been able to obtain a proper spore print or syringe. All is done from obtaining live specimens (Psilocybe caerulescens) that are not mature enough to spore. Not going to delete anything, do not worry.
Thanks for the input ppl.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625251 - 09/27/14 11:00 AM (10 years, 3 months ago) |
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It is possible to take a clone from a wild fruit but you want to take tissue from inside the stem and put it on agar. This process should be done in a still air box using a flame sterilised blade. You should look up some proper agar recipes and preparation methods. Also look into proper sterile procedures.
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ghiajake
Myco-Viking


Registered: 01/10/13
Posts: 3,957
Loc: Indiana
Last seen: 1 day, 20 hours
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625277 - 09/27/14 11:06 AM (10 years, 3 months ago) |
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Quote:
yisroel said:

So, the method you used is wrong like everyone said, and you do need to do much more research at this stage before your practice stage. One thing that is interesting to me is that this actually looks like decent fungal mycelium and not mold or bacteria. I am 100% sure it is contaminated due to your method, but this looks too thick to be mold. I haven't grown myc of that species yet, but it looks healthy (even if not completely clean).
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Trying cultivation from spores, need input. (photos inside) [Re: MudaFuka]
#20625282 - 09/27/14 11:06 AM (10 years, 3 months ago) |
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you need one of these a still air box. use the search function tick the box that says trusted cultivators, search for posts newer than 3 years.
Quote:
PussyFart said: I never clean my room before working in my SAB.......I also don't vaccuum, spray any air cleaners, or use face masks(but you should lol).....all you need is still air, inside and outside the box.
Wear gloves, spray everything down with 70%, alcohol(not 80%), flame your tools.
I use a 5-10% bleach water mixture in the bottom of my SAB, and I spray the walls down with with alcohol.
Works like a charm.
cleaning and isolating on agar
also watch the "let's grow mushroom" videos
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Trying cultivation from spores, need input. (photos inside) [Re: ghiajake]
#20625288 - 09/27/14 11:08 AM (10 years, 3 months ago) |
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Quote:
ghiajake said: One thing that is interesting to me is that this actually looks like decent fungal mycelium and not mold or bacteria
there's cube mycelium in there but it's completely infested with bacteria.
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tripdawg420
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Re: Trying cultivation from spores, need input. (photos inside) [Re: bodhisatta]
#20625321 - 09/27/14 11:13 AM (10 years, 3 months ago) |
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good try tho clean up a lil and you got it
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ghiajake
Myco-Viking


Registered: 01/10/13
Posts: 3,957
Loc: Indiana
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625348 - 09/27/14 11:20 AM (10 years, 3 months ago) |
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Quote:
yisroel said: All is done from obtaining live specimens (Psilocybe caerulescens) that are not mature enough to spore.
OP said they aren't cubes. Either way, it shows that even without any real knowledge or sterile procedures one can germinate spores enough to do an outside grow with.
yisroel don't let any of us discourage you by any means, we're merely giving constructive criticism. Some folks might give you a bunch of shit, but I can honestly say you had better results with those jars than I did when I first started. Must mean you have a knack for it, can't wait to see how you progress after doing appropriate research and follow sterile procedures.
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yisroel
Stranger
Registered: 09/27/14
Posts: 4
Last seen: 10 years, 2 months
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Re: Trying cultivation from spores, need input. (photos inside) [Re: ghiajake]
#20625375 - 09/27/14 11:27 AM (10 years, 3 months ago) |
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Quote:
So, the method you used is wrong like everyone said, and you do need to do much more research at this stage before your practice stage. One thing that is interesting to me is that this actually looks like decent fungal mycelium and not mold or bacteria. I am 100% sure it is contaminated due to your method, but this looks too thick to be mold. I haven't grown myc of that species yet, but it looks healthy (even if not completely clean).
It was kind of a surprise to me. I filled the jars 3/4. First two days I saw nothing at all, then I forgot about them, this sat its a week since inoculation, and the empty space is filled with this cushion, its quite thick.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Trying cultivation from spores, need input. (photos inside) [Re: yisroel]
#20625435 - 09/27/14 11:41 AM (10 years, 3 months ago) |
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the fact that the jars looks so thick and the space between the grains is filled is the indicator of bacterial contamination, I've actually never seen jars that have looked that bad before, if you bury that outside it may yield something but any chances of an indoor grow are gone.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Trying cultivation from spores, need input. (photos inside) [Re: bodhisatta]
#20625480 - 09/27/14 11:53 AM (10 years, 3 months ago) |
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The only time I've had jars look like that Was my horribly contaminated enoki jars that had both bacteria and mold. They looked just like that right before they changed colour from white to green, yellow and brown.
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ghiajake
Myco-Viking


Registered: 01/10/13
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Re: Trying cultivation from spores, need input. (photos inside) [Re: MudaFuka]
#20625754 - 09/27/14 12:57 PM (10 years, 3 months ago) |
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Quote:
bodhisatta said: if you bury that outside it may yield something but any chances of an indoor grow are gone.
Oh, I agree totally! Just sayin'. My first round of jars was completely green and no white within a week.
Quote:
MudaFuka said: The only time I've had jars look like that Was my horribly contaminated enoki jars that had both bacteria and mold. They looked just like that right before they changed colour from white to green, yellow and brown.
I have had jars that had good mycelium with bacterial contamination that look like that before they start getting yellow and "soupy" from the metabolites, but at a week of 84F temps they would have already turned green if they were full of trich. In the pic of the 4 jars if you look at the top left side of the far right jar, that looks like trich to me. But the rest doesn't.
OP, before tossing them crack the lids open and smell them. Worst case scenario you learn the different smells of different contaminations. Or save them for another week so you can see how the contams develop.
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cronicr



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Re: Trying cultivation from spores, need input. (photos inside) [Re: ghiajake]
#20626044 - 09/27/14 02:00 PM (10 years, 3 months ago) |
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keep plucking away at it you'll get there
-------------------- It doesn't matter what i think of you...all that matters is clean spawn
I'm tired do me a favor
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ghiajake
Myco-Viking


Registered: 01/10/13
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Re: Trying cultivation from spores, need input. (photos inside) [Re: cronicr]
#20626091 - 09/27/14 02:12 PM (10 years, 3 months ago) |
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Quote:
cronicr said: keep plucking away at it you'll get there
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