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InvisiblePastywhyteMDiscord
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26792888 - 06/29/20 06:25 PM (3 years, 6 months ago)

I’m dealing with a yeasty vendor syringe myself. Yeast is growing pretty slow though so I’m thinking it will be pretty easy to grab some clean myc in a day or two.


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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: Pastywhyte]
    #26794607 - 06/30/20 01:44 PM (3 years, 6 months ago)

Quote:

I’m dealing with a yeasty vendor syringe myself. Yeast is growing pretty slow though so I’m thinking it will be pretty easy to grab some clean myc in a day or two.




Great! Please share some pictures of your agar, would love compare the initial growth to what I got on my plates.
What I experienced with the yeast on my plates is that it initially grows very quickly and then just stops growing. It does seem that it really inhibits germination when close by spores.


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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26794642 - 06/30/20 02:05 PM (3 years, 6 months ago)

Agar update!

So I am pretty sure the plate I thought to be mycelium is in actuality mold :puke:

1st observation)
It is 3 days since I transferred this growth from A4 plate and it grows way too quickly. The three pieces have now covered the whole plate:


2nd observation) In the following picture.
To the right: A 7 month old plate of king oyster mycelium (healthy culture that I have fruited).
At the bottom: The growth that I now believe to be mold. It is grey compared to the king oyster mycelium.
At the top: The bacteria infused growth that I now believe to be mycelium.


It's sad that I am most likely growing mold here.
However this made me shift my focus to my other transfers. They grow much slower and also the hyphae seem whiter and more condensed, less airborne and seems to grow in circular sectors. Even when they are growing together with bacteria.



Also I found another plate, A1 to be exhibiting the same kind of growth as those I transferred. But much cleaner looking. Doesn't seem like it has bacteria. Has many circular sectors.


So yeah I am doing more transfers today. But I will stop propagating what I now believe to be mold.


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OfflineVernsMB
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26794677 - 06/30/20 02:24 PM (3 years, 6 months ago)

if you use a stainless straw you can make nice small circular transfers.  Place agar inverted so mycellium is sandwiched and if you have healthy mycellium it will quickly out grow contaims onto clean agar.  Transfer again and then you should see contaim free cultures, assuming no airborne critters make it into your petris.

I made mistake of leaving window open morning before I did a large volume of transfers.  Even with SAB and misting with Alcohol I managed to pickup bacteria/yeast in many of my petris...Starting to suspect it was from supplier as infections appear where mycellium liquid was injected.


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OfflineC12ShroomMan
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26794779 - 06/30/20 02:58 PM (3 years, 6 months ago)

Quote:

krock said:
Agar update!

So I am pretty sure the plate I thought to be mycelium is in actuality mold :puke:

1st observation)
It is 3 days since I transferred this growth from A4 plate and it grows way too quickly. The three pieces have now covered the whole plate:






I wouldn't call that mold until the spores show up in the middle.
All my mycelium has grown faster than mold.
1 transfer per plate would of made this easier :smile:


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InvisiblePastywhyteMDiscord
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: C12ShroomMan]
    #26794818 - 06/30/20 03:16 PM (3 years, 6 months ago)

I’m not convinced that first plate is mold. However if it is cubes they are weak and lack vigor.


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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: Pastywhyte]
    #26794904 - 06/30/20 04:04 PM (3 years, 6 months ago)

Haha, now I doubt myself again.

I just came back from transferring pieces from the different smaller growth areas. Used a small piece for each plate.

I didn't transfer anything from the "mold" plate. I will keep it around and see if it pins. Maybe it is in fact not mold. I hope so. Because it's certainly much faster than the other myc-like grow areas.


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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26796337 - 07/01/20 06:33 AM (3 years, 6 months ago)

Update on the first transfers


A4T1 Day 4:



B2T1 (small growth transfer) Day 5:



B2T1 (big growth transfer) Day 5:



I still think the B2 transfers look more like mycelium than the A4 transfers. I transferred a piece from the A4T1 plate to a new plate today so we can inspect the growth more closely in a few days.


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Offlineredhandmat
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock] * 1
    #26796786 - 07/01/20 11:02 AM (3 years, 6 months ago)

You are letting the plates grow too much before transferring, in the phase you're at its good to be ready to transfer asap before other things than cube (if it is cube) get the chance to develop. Transfer as soon as you have enough to do so. Keep the original plates if possible so you may know if its mold (if it changes color etc)


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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: redhandmat]
    #26797035 - 07/01/20 01:35 PM (3 years, 6 months ago)

Quote:

You are letting the plates grow too much before transferring



You're right. I let the A4T1 plate grow too much. Mostly because I was unsure if it is mold or not.
For the B2T1 plates I couldn't transfer them before yesterday as the hyphae growing past the bacteria patch was not big enough to cut at my current skill level. I remember the suggestion to use hot pour or sandwich method, but seeing as the hyphae started outgrowing the bacteria I think it was better to wait a bit.

Quote:

Transfer as soon as you have enough to do so. Keep the original plates if possible so you may know if its mold



I didn't think about that! It makes perfect sense to inspect the parent plate for changes instead of the newly transferred one.

Again thank you for the advice, it helps a lot!


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InvisiblePastywhyteMDiscord
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26797137 - 07/01/20 02:19 PM (3 years, 6 months ago)

Not the best pic but you can see the mycelium popping up in the centre of the yeast colony. I’m going to try and grab a piece of it. This yeast is really slow growing in these temps so the cubes should able to be isolated.



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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: Pastywhyte]
    #26797169 - 07/01/20 02:34 PM (3 years, 6 months ago)

Thank you for posting your yeasty agar! How did you apply the spore solution? Streak or drop?

I find it interesting that the mycelium is able to grow from the center of the colonies on your plate.

On my plates, the growth that I assume to be mycelium is only germinating when it is spread far enough apart from a yeast colony.
Although my solution is also pretty old, which might explain the low germ rate as well.


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InvisiblePastywhyteMDiscord
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26797369 - 07/01/20 04:12 PM (3 years, 6 months ago)

Most yeasts are pretty weak compared to cubes. Doesn’t really surprise me to be honest. I just dropped right off the syringe. No swab.


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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: Pastywhyte]
    #26798567 - 07/02/20 05:26 AM (3 years, 6 months ago)

Agar update

I've taken pictures of all the plates and their transfers. I see that a lot of the new transfers still have bacteria so I am most likely going to attempt a hot pour soon.


    B2

    Streak plate, day 9:


    T1 plates day 6:


    T2 plates day 2:








    A4

    Streak plate, day 9:


    T1 day 6:


    T2 day 1:





    A1

    Streak plate, day 9:


    T1 day 2:




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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26801159 - 07/03/20 09:52 AM (3 years, 6 months ago)

My agar is just finished cooking in the PC.  Now I am waiting for it to cool down enough to pour.

I will hot pour all the T2 plates in probably 15 min. And also make at least 10 more plates for future transfers.

This is what the T2 plates look like right now:



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InvisiblecoAsTal
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26801297 - 07/03/20 11:22 AM (3 years, 6 months ago)

I'm really impressed-- and amazed you got something out of such a shitty storage situation.

:cheers:


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Invisiblekrock
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26801320 - 07/03/20 11:36 AM (3 years, 6 months ago)

Quote:

I'm really impressed-- and amazed you got something out of such a shitty storage situation.
:cheers:



Thanks man! I just came back from pouring and wrapping 30 plates as well as hot pouring the T2 plates.
Heres hoping for that the myc is cubensis :cheers:


Edited by krock (07/03/20 11:37 AM)


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Offlineredhandmat
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: krock]
    #26802636 - 07/04/20 01:34 AM (3 years, 6 months ago)

I like these! But again I urge you to not wait too long to take T3, and take as small transfers as possible. Hmm one question, why isn't your agar clear? Whats your recipe?


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Offline3some
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: Pastywhyte]
    #26802675 - 07/04/20 01:57 AM (3 years, 6 months ago)

Quote:

Pastywhyte said:
Not the best pic but you can see the mycelium popping up in the centre of the yeast colony. I’m going to try and grab a piece of it. This yeast is really slow growing in these temps so the cubes should able to be isolated.






What temp slows bacteria? Hot or cold? Isn't it summer for you? Where u keep your plates?


--------------------
Ask me about growing yeast, mold and bacteria. I'm very experienced. :tongue2:

The first post any newbie should read even before acquiring spores: Pastywhyte's Principles of mushroom growing for beginners


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Offlineredhandmat
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Re: 4th attempt at cleaning up my yeasty spore solution [Re: 3some]
    #26802693 - 07/04/20 02:14 AM (3 years, 6 months ago)

Quote:

3some said:
Quote:

Pastywhyte said:
Not the best pic but you can see the mycelium popping up in the centre of the yeast colony. I’m going to try and grab a piece of it. This yeast is really slow growing in these temps so the cubes should able to be isolated.






What temp slows bacteria? Hot or cold? Isn't it summer for you? Where u keep your plates?




Cold slows bacteria, and cold enough slows everything down.


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