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Offlinealienascii
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Registered: 05/10/09
Posts: 28
Last seen: 2 months, 25 days
Re: If I build it, will they come? [Re: yogibike]
    #28720330 - 03/30/24 02:46 PM (2 months, 27 days ago)

Hey yogibike,
    Sorry I lost interest when your flow thing wasn't made of cardboard :lol:  Definitely keep a control plate for longer than 8 days to be sure.

    SAB's are solid you don't need a bleach soaked towel or anything like that you probably don't even need to wipe it down but why not.  No need to put your phone in a plastic bag either just clean it with alcohol

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Invisibleyogibike
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Registered: 03/31/18
Posts: 32
Loc: Hot desert climate
Re: If I build it, will they come? [Re: alienascii]
    #28721510 - 03/31/24 03:50 PM (2 months, 26 days ago)

The local PhD who teaches mushroom cultivation is coming over today to look at my plates and advise me on what to do with them.

Since he uses and advises using a Bunsen burner to create an updraft in which to do sterile transfers, and since everyone prefers giving advice to people who *follow* their advice, I bought one -- the one on Amazon that has folding legs and an integrated hose and tank connector. Wow, does this one roar.

I've also dug out my SAB.  This time, I'll line the bottom with a towel soaked in bleach water -- and a wire rack over that.

I'll make some new agar and pour 20 more plates. I might pour half inside the SAB and half with the Bunsen burner.

I'm most concerned about the agar pouring temperature.  Comparing WVTR of LDPE in Ziploc bags versus in Parafilm M, the latter appears to be comparable or lower (0.88) than the former (1-1.5). Given this, I'm not confident that condensation in my plates would have been reduced by using Parafilm.

Last time, I poured the agar at 47C, which is near the low end of the pourable range, and it did start to congeal by the last two of 20 plates. Even so, I had trouble with condensation.  I suspect that the situation was not helped by working in a small bathroom, with no ventilation, made steamy by the 47C water bath for the agar and by the fact that I'd opened my pressure cooker inside it.

My OCD brain has some anxiety about Parafilm because I must handle it and the plates quite a bit in order to cut & apply it.

Edited by yogibike (03/31/24 04:04 PM)

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Invisibleyogibike
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Re: If I build it, will they come? [Re: alienascii]
    #28721518 - 03/31/24 04:00 PM (2 months, 26 days ago)

I still have two plates (marked #7) that don't appear to have grown anything yet, having been poured on 2/25 along with the rest.

I meant to inoculate these on 3/6 along with the others. I marked them #7, which was supposed to be shitake. I should check the shitake syringe to see if it has anything in it. Maybe the vendor forgot to add spores to that one. :smile: If the "shitake" syringe is just sterile water, that would explain why I got no contams in that one. :smile:

I realize that my FFU doesn't give a true laminar flow. And the workable area is pretty small. It's possible that I kept this plate in the workable area and was sloppy with some of the others.

Anything that doesn't happen in that little bathroom is gonna be much more fun for me, even if my contam rate ends up higher than it's been. So I'm feeling good about shelving the FFU for now (even though I think it produced at least two plates that stayed contam-free for 34 days so far) in favor of SAB and Bunsen burner.

Thanks for all your thoughtful replies.

Edited by yogibike (03/31/24 04:21 PM)

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Offlinealienascii
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Registered: 05/10/09
Posts: 28
Last seen: 2 months, 25 days
Re: If I build it, will they come? [Re: yogibike]
    #28721549 - 03/31/24 04:32 PM (2 months, 26 days ago)

Quote:

yogibike said:
Thanks for all your thoughtful replies.




Props for being a gentleman, I have no idea what's going on.

so-

Use your SAB till you get a proper flow hood

Do no-pour plates and open the foil in your SAB you can't fuck it up.

The bunsen burner method is wack, in my humble opinion.

You're way overthinking it just use clingwrap

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Invisibleyogibike
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Posts: 32
Loc: Hot desert climate
Re: If I build it, will they come? [Re: alienascii]
    #28721705 - 03/31/24 06:13 PM (2 months, 26 days ago)

Here's a plate from a syringe of ATL#7 Galindoi, one of the sclerotia-bearing Psilocybes. Does it seem like it might be okay? From here, would you do a transfer to agar for strain isolation -- or start over?

I guess the fact that it's fully colonized means that I can't tell which strain won the "race" toward the edge. Also, if it was like most of the others that day, I didn't manage to get the spores into the middle of the plate anyway, and then the heavy condensation may have carried them all over the place.

I guess I wouldn't mind trying a no-pour agar TEK, but I have 100 more of these ready to noc up!


What would you do with this?
You may choose only one


Votes accepted from (03/31/24 06:13 PM) to (No end specified)
You must vote before you can view the results of this poll


Edited by yogibike (03/31/24 08:38 PM)

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Invisibleyogibike
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Re: If I build it, will they come? [Re: yogibike]
    #28721786 - 03/31/24 08:08 PM (2 months, 26 days ago)

Now I've taken photos of the top surfaces of the agar plates (those of the 20 that had anything growing on them). I put a light on top of my SAB and shot from about 45 degrees above horizontal.

My camera's autofocus had trouble focusing on the especially fluffy white stuff -- like, fluffy enough to touch the lid -- which I'm worried may be contam.

Even at full resolution, it's not easy to see what's going on, but I can't attach them all here, due to Shroomery's weekly & lifetime upload limits. To upload all my images, I'd need to become a supporter, for which the subscription requires a card, which is linked to my real name, unless... Maybe it will accept a prepaid debit card? Even then, now that I think about it (darn my damaged brain!) one should probably use a VPN or Tor to create a new email address just to link with a Shroomery account on which one is posting photos about his attempt to grow psilocybin mushrooms.

Edited by yogibike (03/31/24 08:25 PM)

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OfflineCocaineBuffet
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Re: If I build it, will they come? [Re: yogibike]
    #28721799 - 03/31/24 08:29 PM (2 months, 26 days ago)

Nothing worth transferring from on that plate

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Invisibleyogibike
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Loc: Hot desert climate
Re: If I build it, will they come? [Re: yogibike]
    #28721800 - 03/31/24 08:29 PM (2 months, 26 days ago)

Here's a plate of landscape black morel:

Edited by yogibike (03/31/24 08:39 PM)

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Invisibleyogibike
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Re: If I build it, will they come? [Re: yogibike]
    #28721802 - 03/31/24 08:32 PM (2 months, 26 days ago)

Here's a plate of yellow oyster:

Edited by yogibike (03/31/24 08:39 PM)

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Invisibleyogibike
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Re: If I build it, will they come? [Re: yogibike]
    #28721809 - 03/31/24 08:36 PM (2 months, 26 days ago)

Here's a plate of P. tampensis.

Edited by yogibike (03/31/24 08:39 PM)

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Invisibleyogibike
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Re: If I build it, will they come? [Re: CocaineBuffet]
    #28721853 - 03/31/24 09:02 PM (2 months, 26 days ago)

Quote:

CocaineBuffet said:
Nothing worth transferring from on that plate




Thank you. That was the ATL#7 Galindoi. What do you think of the next three I've posted, below your reply?

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OfflineCocaineBuffet
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Re: If I build it, will they come? [Re: yogibike]
    #28721865 - 03/31/24 09:09 PM (2 months, 26 days ago)

I am not experienced in gourmets so I cant speak on the morel or oyster.

Tamp looks way too wet for an agar plate but I think some of the more rhizo growth would warrant a transfer.

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Offlinealienascii
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Registered: 05/10/09
Posts: 28
Last seen: 2 months, 25 days
Re: If I build it, will they come? [Re: CocaineBuffet]
    #28722255 - 04/01/24 09:30 AM (2 months, 26 days ago)

x2 good job on the pics

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Invisibleyogibike
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Registered: 03/31/18
Posts: 32
Loc: Hot desert climate
Re: If I build it, will they come? [Re: alienascii]
    #28722308 - 04/01/24 10:33 AM (2 months, 25 days ago)

Quote:

alienascii said:
x2 good job on the pics




Thank you! I can post more, especially after I delete some of the previous, less-useful ones. I've realized that Shroomery downscales them quite a bit, so there's room.

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