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Offline3rdEyeVision
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Registered: 07/18/19
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Agar contam help/advice needed
    #26534629 - 03/14/20 11:49 AM (3 years, 10 months ago)

Hey everyone- I attempted my first clone to agar run to learn the steps using pastywhyte's easy agar tek. While a couple of the containers looked off from the beginning, several looked good and the clones progressed properly, but then the clone containers started to develop this off color underneath the surface mycelium. I searched online for images but didn't see anything similar. I took the clones from the inside stem after tearing apart working in the SAB. These look bad to me now but I was hoping to get confirmation from experienced members. Also, should I have attempted to transfer these to new plates at a very early stage before the darker stuff appeared?  Thanks






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Offlinemetalcore_dad
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Registered: 02/05/20
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Re: Agar contam help/advice needed [Re: 3rdEyeVision]
    #26540677 - 03/17/20 12:15 PM (3 years, 10 months ago)

Hello friend,


No contam that I can tell. Common contam in SAB work is rhizopus, which can be mistaken for mushroom myc. You'd have to let that grow out however. Rhizopus will climb upward and toward the edges of the container.

Gas and metabolites likely diffusing into the agar and causing color change. Best bet? Always smell it. Your clones look fine, and again the big things I would look out for in agar work are smells, signs of rhizopus (transparent fluffy mycelium, smells like sour/alcohol). If you are truly worried, take an edge and transfer. Also upgrade to actual sterilized dishes, use malt extract agar.....you spend more money buying the stupid ziploc things and you have to twist a cap off vs. just removing a top with 2 fingers using an actual dish.

its 12$/30 sterilized agar plates -- go get some.

Peace


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Offlineredhandmat
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Registered: 05/09/19
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Re: Agar contam help/advice needed [Re: metalcore_dad]
    #26543805 - 03/19/20 02:04 AM (3 years, 10 months ago)

Hey dude, my agar dishes with green colorant got the exact same color as yours beneath the mycelium. Its totally normal.


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Offline3rdEyeVision
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Re: Agar contam help/advice needed [Re: metalcore_dad]
    #26586277 - 04/08/20 12:39 PM (3 years, 9 months ago)

Quote:

metalcore_dad said:
Hello friend,


No contam that I can tell. Common contam in SAB work is rhizopus, which can be mistaken for mushroom myc. You'd have to let that grow out however. Rhizopus will climb upward and toward the edges of the container.

Gas and metabolites likely diffusing into the agar and causing color change. Best bet? Always smell it. Your clones look fine, and again the big things I would look out for in agar work are smells, signs of rhizopus (transparent fluffy mycelium, smells like sour/alcohol). If you are truly worried, take an edge and transfer. Also upgrade to actual sterilized dishes, use malt extract agar.....you spend more money buying the stupid ziploc things and you have to twist a cap off vs. just removing a top with 2 fingers using an actual dish.

its 12$/30 sterilized agar plates -- go get some.

Peace




Thanks for the guidance and you're right, they were OK. Being a first time agar run, I wasn't sure about that discoloring but I let these ride when I wasn't seeing anything off on the surface. I transferred into new dishes recently and so far so good. I also tried out a couple on grain when I ran a batch of jars and the colonizing is moving along super fast and clean.

Thanks for the tips on Rhizopus as well. Now that I have an idea of how things work, I am looking forward to moving to the real plates to learn more about the process.

-Funny, I actually jumped on to update my post and noticed that some people responded so I greatly appreciate the help!


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Offline3rdEyeVision
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Re: Agar contam help/advice needed [Re: redhandmat]
    #26586317 - 04/08/20 12:57 PM (3 years, 9 months ago)

Quote:

redhandmat said:
Hey dude, my agar dishes with green colorant got the exact same color as yours beneath the mycelium. Its totally normal.




Hey man! Thank you for the help and confirmation. Being new to agar, I was thrown off by the color change but decided to trust what I was seeing on the surface and let them keep going. I know that I am in for more head scratching and questions with agar but getting a run or two under the belt has been super helpful.

I just started last summer with a simple pf tek run and was totally hooked after that!

Cheers!


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