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veryannoyed
Stranger

Registered: 07/29/21
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Why do my agar plates end up with these defects? 1
#27655616 - 02/11/22 07:53 PM (2 years, 3 months ago) |
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So I’ve been making agar plates for a few months now and have had great success with it. Occasionally my plates will show signs of me not doing something right. I have a good sterile technique and pour them in a still air box but like I said occasionally my jars have the same kind of defects showing as in the pictures. What would cause this? Am I pouring it too hot or cold? Is the agar or malt not dissolving completely? I really do not know what I am doing wrong and any help from you guys would be great. https://
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verum subsequentis
seeker of truth



Registered: 03/22/16
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Re: Why do my agar plates end up with these defects? [Re: veryannoyed] 1
#27655913 - 02/12/22 01:15 AM (2 years, 3 months ago) |
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I hope this is a joke. No harm intended if not. It's just that you posted it in advanced and .... well. It's clearly not. Your plates are as fucked as fucked can be.
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DERRAYLD
Constructus

Registered: 05/13/02
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Loc: South Africa
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Those are all bacterial, your technique is poor.
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hongito33
Lord of the detritivores


Registered: 10/22/18
Posts: 144
Loc: in your head
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Re: Why do my agar plates end up with these defects? [Re: veryannoyed] 2
#27656870 - 02/12/22 09:56 PM (2 years, 3 months ago) |
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Quote:
veryannoyed said: So I’ve been making agar plates for a few months now and have had great success with it. Occasionally my plates will show signs of me not doing something right. I have a good sterile technique and pour them in a still air box but like I said occasionally my jars have the same kind of defects showing as in the pictures. What would cause this? Am I pouring it too hot or cold? Is the agar or malt not dissolving completely? I really do not know what I am doing wrong and any help from you guys would be great. https://
Ok, probably someone already pointed the obvious, whether you are an advance mycologist or not, your sterile technique needs some brushing up. What you have there my friend is bacterial infection that has been created by either a contaminated environment or your technique need more work. I do not mean to criticize you, many of us have been there and no one gets is right the first few times. First things first:
1.Are you doing your work in a still air box or laminar flow hood? 2.When you sterilize your agar, what is your sterilization times? 3.are your plates Sterilized? 4.When you pour your plates do they get condensation and water pools on the agar?
if you do not know the answers to these questions i would suggest you post in the Agar envy section of mushroom cultivation.
regards,
Hongito33
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Bobbins
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Re: Why do my agar plates end up with these defects? [Re: hongito33] 1
#27662084 - 02/17/22 01:37 AM (2 years, 2 months ago) |
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That's a lot of bacteria around the periphery, I'm guessing it's getting in while the plates are stored/handled untaped.
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veryannoyed
Stranger

Registered: 07/29/21
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Re: Why do my agar plates end up with these defects? [Re: hongito33] 1
#27663505 - 02/18/22 05:26 AM (2 years, 2 months ago) |
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Thanks for the reply. Obviously I know they are contaminated. I just don’t know when it is happening. I showed 4 that happened to have been inoculated already but the same types are on plates I have not used.
I am using a still air box, I sterilize the agar in a pressure cooker. Once the pressure cooker reaches 15psi I set a timer for 20 minutes. I also have tape that indicates if I have reach sterilization temperatures. I then move my flask to the still air box and check the temp with a temp gun. Once the agar reaches 55-60 degrees Celsius I pour my plates. When I purchased the plates they say they are sterile besides that I wouldn’t know. I am trusting the seller at that point. When I pour the plates I have never seen water pool up, they do get some condensation but never pooled water. I have sealed the SAB and let the plates sit for about 30 min before so they can dry out.
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DERRAYLD
Constructus

Registered: 05/13/02
Posts: 10,170
Loc: South Africa
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Re: Why do my agar plates end up with these defects? [Re: veryannoyed] 1
#27663523 - 02/18/22 06:16 AM (2 years, 2 months ago) |
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If it's happening on unused plates it points to your pc cycle or the storage of the plates after.
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coversall
إِنْ شَاءَ ٱللَهُ



Registered: 06/06/20
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Re: Why do my agar plates end up with these defects? [Re: DERRAYLD] 1
#27664694 - 02/19/22 03:29 AM (2 years, 2 months ago) |
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Are you making sure the not hoover your hand over the plate at all while pouring? You want to hold the media bottle near the base and turn the wrist so that your hand is never directly over the plate while pouring.
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HotMess
i'm your buddy
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Re: Why do my agar plates end up with these defects? [Re: coversall] 1
#27670791 - 02/23/22 07:24 PM (2 years, 2 months ago) |
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the boogery bacteria at the edges of some plates, i have noticed i would often get that if i had a lot of condensation in my plate, enough to pool or make large drops in the lid when stored upside down. then when i turn the plate over to work the drops get on the agar and cause all that bacteria. not sure why this happens, i must be getting bacteria on the inside of the lid somehow but i never touch it.
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The Seeker Of One
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Re: Why do my agar plates end up with these defects? [Re: veryannoyed] 2
#27671078 - 02/24/22 12:44 AM (2 years, 2 months ago) |
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Try pouring the plates at 47 Deg C.
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The Seeker Of One
Stranger



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Re: Why do my agar plates end up with these defects? [Re: veryannoyed] 1
#27671084 - 02/24/22 12:48 AM (2 years, 2 months ago) |
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Quote:
veryannoyed said: Thanks for the reply. Obviously I know they are contaminated. I just don’t know when it is happening. I showed 4 that happened to have been inoculated already but the same types are on plates I have not used.
I am using a still air box, I sterilize the agar in a pressure cooker. Once the pressure cooker reaches 15psi I set a timer for 20 minutes. I also have tape that indicates if I have reach sterilization temperatures. I then move my flask to the still air box and check the temp with a temp gun. Once the agar reaches 55-60 degrees Celsius I pour my plates. When I purchased the plates they say they are sterile besides that I wouldn’t know. I am trusting the seller at that point. When I pour the plates I have never seen water pool up, they do get some condensation but never pooled water. I have sealed the SAB and let the plates sit for about 30 min before so they can dry out.
Are you letting the plates sit uncovered for 30 Mins ?
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Putrema
1shroomis1shroomer

Registered: 02/20/20
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Re: Why do my agar plates end up with these defects? [Re: The Seeker Of One] 1
#27704361 - 03/22/22 07:59 AM (2 years, 1 month ago) |
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Lol now Im feeling better, because someone posted this in the advanced forum and Im better than youuuu. I did pouring in a crinkled cardboad SAB with (at least partially) better results. But there is a big wuuuuuut factor that I cant estimate. Why are some plates so much better than other? Yeah, its exciting. To narrow down the mistakes but to respect the unknown factor also. How big is the heater turbulence in there? Better throw a blanket in front of the door (who knows what kind of turbulences lurk there)? Good luck.
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Rotnpins
🤮 Rotten-Pins 🍄



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Re: Why do my agar plates end up with these defects? [Re: Putrema] 1
#27705435 - 03/23/22 06:00 AM (2 years, 1 month ago) |
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primordialkraken
merciless


Registered: 08/15/18
Posts: 112
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Re: Why do my agar plates end up with these defects? *DELETED* [Re: veryannoyed] 1
#27724934 - 04/07/22 07:30 PM (2 years, 1 month ago) |
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Post deleted by primordialkraken
Reason for deletion: .
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Shunyata
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Re: Why do my agar plates end up with these defects? [Re: primordialkraken] 1
#27726073 - 04/08/22 05:57 PM (2 years, 1 month ago) |
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Try using different dishes or souffle cups and see if you have the same problem. I rarely ever see any contams using these plastic cups and it's all I use. https://tinyurl.com/3jyh7vej
If you're sure i'ts not the PC then maybe there is a lot of dust in the air and it's getting in your SAB?
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TheConfluence
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Registered: 12/13/21
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Re: Why do my agar plates end up with these defects? [Re: Shunyata] 1
#27735676 - 04/15/22 09:56 AM (2 years, 1 month ago) |
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Bacteria collected in the condensation that formed in the plate due to improper cooling. A HEPA or an SAB will pretty much completely solve the issue.
Is it bacillus?
-------------------- When two people dream the same dream, it ceases to be an illusion. Philip K. Dick Lies, Inc. (1984)
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Nillion
Nobody

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Re: Why do my agar plates end up with these defects? [Re: TheConfluence] 1
#27737642 - 04/16/22 06:04 PM (2 years, 1 month ago) |
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Quote:
TheConfluence said: Bacteria collected in the condensation that formed in the plate due to improper cooling. A HEPA or an SAB will pretty much completely solve the issue.
Is it bacillus?
He mentioned using an SAB, but it isn't clear what the vector is yet. A different SAB might be required.
A lot of cultivators use agar with anti-bacterial properties to avoid issues like this. It is a shortcut that will not replace clean technique but it is an option nonetheless.
Edited by Nillion (04/16/22 06:05 PM)
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MycoWeek
Elementary Student



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Loc: USA
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Re: Why do my agar plates end up with these defects? [Re: Nillion] 1
#27738691 - 04/17/22 12:57 PM (2 years, 1 month ago) |
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almost all bacteria colonies are around the edge, I am suspecting it's "sucked in" after the lid is closed.
did you wrap them with parafilm? and were they wrapped before or after agar is completely cooled?
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