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OfflinePhilberto
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Registered: 08/31/22
Posts: 255
Last seen: 1 year, 6 months
Hoping I'm on the right track.
    #28009809 - 10/21/22 06:43 PM (2 years, 2 months ago)

I was learning from a friend who is potentially a beginner. He had me purchase a spore syringe and put it directly into prepared seed, instead of putting it on to agar first. I also put some of the spore syringe into a solution to create my own liquid culture. It appeared that mycelium began to grow normally in both.
I noticed in the liquid culture after a couple of weeks there was a very minor spot that was a tiny bit darker. (I'm colorblind, and I assumed the worst)  I figured that perhaps it was contamination so started the process of making agar plates.
Of the 16 plates I created, I put the liquid culture I had created into eight of them.
They got white fluffy growths on them soon after, however one developed a green spot and clearly had a contamination. I set it aside just to see how it would develop. I then noticed five of my unused plates developed a dot of contamination within 5 or 6 days, most likely because I carried them around the house without them being tape sealed.
The other unused plates are still perfectly clean.

So logic.....  Since seven of the eight inoculated plates remain uncontaminated, does that not suggest that my liquid culture is actually good?  I'm assuming that my handling of the agar plates before inoculating them was what caused those few to have a spot of contamination. 
I have since taped all of the containers so nothing can enter, and placed all of the unused agar dishes into a Tupperware container. 

Theoretically I wouldn't have seven good examples of mycelium on agar coming from contaminated liquid culture.... Right?

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OfflinePadaone
Mycophage

Registered: 07/03/22
Posts: 68
Last seen: 3 months, 27 days
Re: Hoping I'm on the right track. [Re: Philberto]
    #28009816 - 10/21/22 06:50 PM (2 years, 2 months ago)

This will be a whole lot easier to determine with pics.

How did you prep your agar? Are you using a SAB. Way back in the day when I tried my first (and only at that time) bulk grow spores to grain never worked well for me. Agar is the way, you'll surpass your friends cult skills in no time with that advice they're giving.

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OfflinePhilberto
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Registered: 08/31/22
Posts: 255
Last seen: 1 year, 6 months
Re: Hoping I'm on the right track. [Re: Padaone]
    #28009829 - 10/21/22 06:57 PM (2 years, 2 months ago)

https://files.shroomery.org/files/22-42/640016225-received_528791202413725.jpg

This is a picture of what I believe to be healthy stuff. I'm just guessing that I couldn't have this be healthy if my liquid culture was bad.

I copied the PGT YouTube version of agar.

Aaaannnddd I have no idea what an SAB is.

I didn't realize that cultivation on agar was an advantage. I guess I will do that from now on.

EDIT....  I googled SAB haha. No I made a big flow hood. 

I agree that I might end up surpassing him. It does seem like he complains about throwing away a lot of contaminated grain.  I just tried to set about it being as professional as I could to minimize the risks.

I believe that the fact that I was moving these plates around the house without tape was potentially my issue. The lids don't seem to fit very well on the petri dishes and I feel like some air could move in and out of there.  But plenty of them are still clean

Edited by Philberto (10/21/22 07:08 PM)

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OfflineJaksavage
Mycodelic
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Registered: 11/19/21
Posts: 548
Loc: Left coast
Last seen: 1 year, 5 months
Re: Hoping I'm on the right track. [Re: Philberto]
    #28009850 - 10/21/22 07:22 PM (2 years, 2 months ago)

Welcome to the right place.
You are on the right track.
Check out PF tek, it's a great way to get started.

Do some study and share what you are doing and you will recieve great help here.

:goodluck:


--------------------
:bigbang:

:takingnotes:


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Invisiblemilkboy
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Registered: 05/01/21
Posts: 2,524
Trusted Cultivator
Re: Hoping I'm on the right track. [Re: Jaksavage]
    #28009856 - 10/21/22 07:25 PM (2 years, 2 months ago)

Also, dont feed the animals


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OfflinePhilberto
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Registered: 08/31/22
Posts: 255
Last seen: 1 year, 6 months
Re: Hoping I'm on the right track. [Re: Jaksavage]
    #28009858 - 10/21/22 07:26 PM (2 years, 2 months ago)

I'm guessing I'm extra stupid. What's PF tek?

And can anybody explain what is meant by isolating good strains from your agar plates? Does that just mean picking the strongest fluffiest examples on there?

On my plate that had a small amount of contamination, I did section out a healthy part and put it on a plate just to see if it would grow clean. This stuff is fascinating

Edited by Philberto (10/21/22 07:28 PM)

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OfflineJaksavage
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Re: Hoping I'm on the right track. [Re: Philberto]
    #28009860 - 10/21/22 07:28 PM (2 years, 2 months ago)

Search feature is your friend...

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OfflinePhilberto
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Re: Hoping I'm on the right track. [Re: Jaksavage]
    #28009862 - 10/21/22 07:29 PM (2 years, 2 months ago)

Fair point

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InvisibleMethodicalMystical
Just a normal living being
I'm a teapot User Gallery

Registered: 07/16/17
Posts: 250
Loc: PNW
Re: Hoping I'm on the right track. [Re: Philberto]
    #28009869 - 10/21/22 07:36 PM (2 years, 2 months ago)

Quote:

Philberto said:
https://files.shroomery.org/files/22-42/640016225-received_528791202413725.jpg

This is a picture of what I believe to be healthy stuff. I'm just guessing that I couldn't have this be healthy if my liquid culture was bad.




That plate is about a week old, right? Personally, I'm concerned with how wispy and patchy it looks.

An important thing to keep in mind is that spores are inherently unclean. When spores are taken from a mushroom, nasties such as mold spores and bacteria can hitch a ride from the place it was fruited. Consider your LC compromised and work on clean agar technique before making another.

A lot of your questions can be answered by checking out the Getting Started master post. There you will find links that explain PF tek, agar, and a lot more.


--------------------
I know I'm a fool, but that's why I keep trying to learn.
:peace::heart::mushroom2:

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OfflinePhilberto
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Registered: 08/31/22
Posts: 255
Last seen: 1 year, 6 months
Re: Hoping I'm on the right track. [Re: MethodicalMystical]
    #28009872 - 10/21/22 07:43 PM (2 years, 2 months ago)

Thanks for the input, and just a possibility here....
I discovered after the fact that I was supposed to put one drop from the spore syringe onto the plate. I literally flooded the plate. So I think perhaps it looks wispy and patchy because it is growing from many different locations where it should just be one spot that was growing. (ok... All theory and guesswork)

And yes, they are about a week old. I discovered today however that the temperature where they are kept is perhaps not sufficient. They were at room temperature in a basement, around 65°
I just moved them to a location that stays at 72°

Edited by Philberto (10/21/22 07:44 PM)

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InvisibleMethodicalMystical
Just a normal living being
I'm a teapot User Gallery

Registered: 07/16/17
Posts: 250
Loc: PNW
Re: Hoping I'm on the right track. [Re: Philberto]
    #28009918 - 10/21/22 08:12 PM (2 years, 2 months ago)

That's one thing that happens when you flood a plate with spore water, yes. But also, more spore water means more of those ride-along nasties I mentioned. Your myc is probably competing for space on the plate with those other things.

As far as temperature goes, 65-75 is fine. Just try not to go above 80, as some contams start to get an advantage. 72 should be a nice medium.


--------------------
I know I'm a fool, but that's why I keep trying to learn.
:peace::heart::mushroom2:

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OfflinePhilberto
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Registered: 08/31/22
Posts: 255
Last seen: 1 year, 6 months
Re: Hoping I'm on the right track. [Re: MethodicalMystical]
    #28009940 - 10/21/22 08:39 PM (2 years, 2 months ago)

Thank you for the help. I think I am literally pissing in the wind here despite studying for weeks at a time.

The plates that I flooded obviously are starting to grow in multiple areas, but it does all look clean.
Something I did a few days ago is section out just a small bump of mycelium and put it onto fresh agar by itself. I suppose I should just let those grow out and see how they do and toss these original plates. Does that seem reasonable?
If they grow clean and healthy can I just go straight to bulk media? Or do I need to aim at sectioning and getting a monoculture?
(Genuinely struggling to understand why I need a monoculture, but perhaps it's more important than I realize)

Edited by Philberto (10/21/22 08:40 PM)

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InvisibleMethodicalMystical
Just a normal living being
I'm a teapot User Gallery

Registered: 07/16/17
Posts: 250
Loc: PNW
Re: Hoping I'm on the right track. [Re: Philberto]
    #28010106 - 10/21/22 11:29 PM (2 years, 2 months ago)

You don't need a monoculture. That's something more experienced cultivators do when they find a fruit they like and try to keep a copy of its genes. For now, just focus on having clean and healthy myc. It can take a couple transfers, but you're already on your way there :grin:


--------------------
I know I'm a fool, but that's why I keep trying to learn.
:peace::heart::mushroom2:

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OfflinePhilberto
Actual tosspot
Registered: 08/31/22
Posts: 255
Last seen: 1 year, 6 months
Re: Hoping I'm on the right track. [Re: MethodicalMystical]
    #28010116 - 10/21/22 11:42 PM (2 years, 2 months ago)

Honestly that is such a relief :-)
So as long as it's clean I can grow it... Right?

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OfflineJaksavage
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Registered: 11/19/21
Posts: 548
Loc: Left coast
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Re: Hoping I'm on the right track. [Re: Philberto]
    #28012669 - 10/23/22 02:40 PM (2 years, 2 months ago)

Yes to the clean is good to grow question.

:goodluck:


--------------------
:bigbang:

:takingnotes:


The Shroomery is the biggest library with the coolest librarians

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OfflinePhilberto
Actual tosspot
Registered: 08/31/22
Posts: 255
Last seen: 1 year, 6 months
Re: Hoping I'm on the right track. [Re: Jaksavage]
    #28012748 - 10/23/22 03:34 PM (2 years, 2 months ago)

Thain Q

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