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OfflineLoriFL
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Advice on how my agar plates are doing
    #28365377 - 06/19/23 11:02 AM (1 year, 6 months ago)

Hey everyone! This is my second time doing agar plates and my first ones looked weird. I am making a still air box today, so this was not done in a completely sterile environment. Two of the plates are samples taken from inside the stalk of 2 harvested mushies and one is just a tiny shroom that was attached to a larger shroom that broke its' veil. Can you tell me if these look like they are going to be healthy or is it another bust? Thank you :smile:



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OfflineScrewup
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Re: Advice on how my agar plates are doing [Re: LoriFL]
    #28365379 - 06/19/23 11:03 AM (1 year, 6 months ago)

Bacteria over riding what myc is coming off those pieces.


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InvisibleChRnZN
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Re: Advice on how my agar plates are doing [Re: LoriFL]
    #28365412 - 06/19/23 11:33 AM (1 year, 6 months ago)

Try spores instead of tissue samples - your cuttings and the little shroom will not do anything on agar. You need spores to germinate and become mycelium, the stuff you used is "dead" in the sense that it will not regenerate itself or produce offspring.

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InvisibleMycolorado
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Re: Advice on how my agar plates are doing [Re: ChRnZN] * 3
    #28365417 - 06/19/23 11:39 AM (1 year, 6 months ago)

Dude, you need to stop with the bad advice.  Tissue culturing is done all the time.

OP - As mentioned, you have a lot of bacteria on those plates which speaks to poor technique.  Plating anything outside of inner tissue is a no go as it’s exposed to open air. 

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OfflineLoriFL
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Re: Advice on how my agar plates are doing [Re: Mycolorado]
    #28365434 - 06/19/23 11:54 AM (1 year, 6 months ago)

Quote:

Mycolorado said:
Dude, you need to stop with the bad advice.  Tissue culturing is done all the time.

OP - As mentioned, you have a lot of bacteria on those plates which speaks to poor technique.  Plating anything outside of inner tissue is a no go as it’s exposed to open air. 




Ok. I figured they were contaminated because they don't look like the images I've seen. I was just testing in open air (I know, that's bad). My SAB will be ready today. I have a lot of beautiful spore prints that I made a few days ago, I think I am going to stick with making spore syringes. All of those supplies should be here today. From what I've read, I can get about 2 10mL syringes out of 1 spore print. Does that sound about right to you?


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InvisibleMycolorado
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Re: Advice on how my agar plates are doing [Re: LoriFL] * 1
    #28365440 - 06/19/23 11:56 AM (1 year, 6 months ago)

If you have agar, there’s no need to make syringes. Just flame sterilize a loop and transfer spores from print to plate.

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OfflineLoriFL
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Re: Advice on how my agar plates are doing [Re: Mycolorado] * 1
    #28365455 - 06/19/23 12:05 PM (1 year, 6 months ago)

Quote:

Mycolorado said:
If you have agar, there’s no need to make syringes. Just flame sterilize a loop and transfer spores from print to plate.




Oh, ok! I will try that today. If it is healthy, I should be able to tell within a few days, yeah? Thank you so much for your help, I appreciate it. :smile:


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InvisibleMycolorado
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Re: Advice on how my agar plates are doing [Re: LoriFL]
    #28365457 - 06/19/23 12:07 PM (1 year, 6 months ago)

Yep.  Fresh spores should germinate within a few days.

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Offlinejohnukguy
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Re: Advice on how my agar plates are doing [Re: LoriFL]
    #28365684 - 06/19/23 02:33 PM (1 year, 6 months ago)

Quote:

LoriFL said:
Hey everyone! This is my second time doing agar plates and my first ones looked weird. I am making a still air box today, so this was not done in a completely sterile environment.




To be clear, nothing about a still air box is sterile. The goal is still air, not a sterile environment. If you are making spore syringes, agar is definitely the way to go, unless you are going for a PF Tek grow.
But don't do that in open air unless you are happy for a high degree of contamination. A still air box by the way is often just a larger size tote with a couple of arm holes cut in it.
The premade ones that can be bought online often aren't very good and are a lot more expensive.


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InvisibleChRnZN
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Re: Advice on how my agar plates are doing [Re: Mycolorado]
    #28366077 - 06/19/23 06:36 PM (1 year, 6 months ago)

Quote:

Mycolorado said:
Tissue culturing is done all the time.




Show your References or prove it yourself.

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InvisibleMycolorado
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Re: Advice on how my agar plates are doing [Re: ChRnZN] * 1
    #28366100 - 06/19/23 06:44 PM (1 year, 6 months ago)

You can’t be serious.  TC is common practice.  People clone fruit and post it on here daily. 
Here’s a tissue sample from a few weeks ago.


Even dried material can be revived.
https://www.shroomery.org/forums/showflat.php/Number/28354871#28354871

Here are dried inoculated grains jumping off after 2 days. It’s not TC per se but illustrates the resilience of fungal tissue and its ability to revive even after being dry for some time. 


Your understanding of cultivation is severely lacking and you should stop posting advice in this area and stick to looking up IDs on the internet.

Edited by Mycolorado (06/19/23 07:01 PM)

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InvisibleBaba Yaga
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Re: Advice on how my agar plates are doing [Re: ChRnZN] * 2
    #28366117 - 06/19/23 06:54 PM (1 year, 6 months ago)

OP, don't get discouraged by the one guy who clearly doesn't know what he is talking about. Next time you try your hand at cloning try it with water agar which is just water and agar with no nutrients. Since there are no nutrients in the agar bacteria will grow very slowly and the mycelium has plenty of time to grow out clean. You can simply cut a tissue sample or take a pin and place it on agar. It can't get any easier than that.

Here are a few examples of tissue and pins I took to water agar, they have not been cleaned or treated in any way.


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OfflineLoriFL
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Re: Advice on how my agar plates are doing [Re: johnukguy]
    #28366672 - 06/20/23 06:17 AM (1 year, 6 months ago)

Quote:

johnukguy said:
Quote:

LoriFL said:
Hey everyone! This is my second time doing agar plates and my first ones looked weird. I am making a still air box today, so this was not done in a completely sterile environment.




To be clear, nothing about a still air box is sterile. The goal is still air, not a sterile environment. If you are making spore syringes, agar is definitely the way to go, unless you are going for a PF Tek grow.
But don't do that in open air unless you are happy for a high degree of contamination. A still air box by the way is often just a larger size tote with a couple of arm holes cut in it.
The premade ones that can be bought online often aren't very good and are a lot more expensive.





I do the Uncle Ben's Tek for the spawn. Nope, won't do that in open air again LOL. I do have the SAB now. I feel like there is still so much to learn. I have 6 monotubs going right now. I have a dedicated bedroom with humidifiers and grow lights 12hrs on, 12hrs off. My very first flush on the first tub was small, but by the 5th flush, the mushies were way bigger. I'm trying to figure out what is lacking so all of them grow big like the 5th flush did.The hygrometer reads between 85-90% all the time, they are tended to regularly throughout the day, so I am at a loss as to why the small mushies pop their veils so quickly. I was thinking it may be the FAE, was thinking of going the computer fan way but I dunno. I have a lot of spore prints but I have not used them to inoculate yet. I still have a few syringes left from one of this site's trusted vendors, I just want to be able to clone my own to save costs. I do 6 uncle ben's bags a week and have been for about 6 weeks. So each week I am inoculating and planting the spawn. I keep the uncle ben's in the bathroom in that bedroom, with the light off and a small humidifier. I haven't had any problems with my spawn bags, they have all came out great. I just get small shrooms that pop their veil too early. Oh, and I do the CVG recipe for substrate. Can you think of what I could be doing wrong?


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OfflineScrewup
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Re: Advice on how my agar plates are doing [Re: ChRnZN]
    #28366684 - 06/20/23 06:36 AM (1 year, 6 months ago)

Quote:

ChRnZN said:
Quote:

Mycolorado said:
Tissue culturing is done all the time.




Show your References or prove it yourself.



Quite confident in your incorrectness. I like it b


Stop using Shit bens and do PFTEK. No one cares about RH. Maintain surface conditions via thousands of micro droplets in the surface. No one needs humidifies or computer fans you’re just making things worse. Spawn PFTEK to bulk or use a water rub and leave it the fuck alone. None of your spawn bags are great I can guaranfuckingtee that since you’re working with shit bens.


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Edited by Screwup (06/20/23 06:38 AM)

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OfflineLoriFL
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Re: Advice on how my agar plates are doing [Re: Screwup]
    #28366716 - 06/20/23 07:13 AM (1 year, 6 months ago)

Quote:

Screwup said:
Quote:

ChRnZN said:
Quote:

Mycolorado said:
Tissue culturing is done all the time.




Show your References or prove it yourself.



Quite confident in your incorrectness. I like it b


Stop using Shit bens and do PFTEK. No one cares about RH. Maintain surface conditions via thousands of micro droplets in the surface. No one needs humidifies or computer fans you’re just making things worse. Spawn PFTEK to bulk or use a water rub and leave it the fuck alone. None of your spawn bags are great I can guaranfuckingtee that since you’re working with shit bens.




You don't have to be rude


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OfflineScrewup
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Re: Advice on how my agar plates are doing [Re: LoriFL]
    #28366721 - 06/20/23 07:17 AM (1 year, 6 months ago)

The first sentence wasn’t for you but the rest was FWIW lol


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