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InvisibleTheDoobskerS
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Registered: 05/22/20
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Having troubles with agar
    #26922933 - 09/07/20 12:10 PM (3 years, 5 months ago)

I've been cloning some of my fruits lately and not really having good luck.

My first time cloning, I cloned 2 different cube strains and got really good growth on almost every single plate. But then I made some transfers from those clones and got this. All of the NC transfer plates look just like this one.






This KSSS plate was looking good to me after the transfer but now it's starting to have inconsistent growth. I was planning to put it to grain in the next couple days, but now idk



Any advice? I don't see how they could all be contaminated, but they don't look right to me.

I'll be filling up like 10 plates with clones later tonight and also gonna try putting pins on a couple plates, so we'll see how those go


Edit: This plate was a clone from this Z-strain mushroom. Definitely some bacterial contamination there, but why do all my clean clones look like this?





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Edited by TheDoobsker (09/07/20 12:31 PM)


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Offlineverum subsequentis
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Registered: 03/22/16
Posts: 8,732
Last seen: 1 year, 8 months
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Re: Having troubles with agar [Re: TheDoobsker]
    #26923129 - 09/07/20 01:48 PM (3 years, 5 months ago)

They may not be clean. The third one almost definitely has some bacteria chilling up in there. I'd strongly suggest pouring plates a week or so before you need to use them so that you can be sure that they are clean before use. Other than that you just need to make another transfer from each and see how they turn out. Always transfer from the fastest growing, sexiest looking and most organized looking portion of a plate.


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InvisibleTheDoobskerS
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Re: Having troubles with agar [Re: verum subsequentis]
    #26923278 - 09/07/20 03:30 PM (3 years, 5 months ago)

Would you suggest making the plates and just leaving them in room temp until using them?
I tried making plates in bulk before but just go so much condensation from storing them in the fridge that I stuck with making 5-10 plates at a time the night before I'll be using them.

Is it common to need to do some transfers to get a nice looking culture?
I was thinking as long a clone plate is clean, the next transfer plate should be able to go to grain


I appreciate the advice :thumbup:


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Offlineverum subsequentis
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Registered: 03/22/16
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Re: Having troubles with agar [Re: TheDoobsker]
    #26923385 - 09/07/20 04:53 PM (3 years, 5 months ago)

There is absolutely no reason to store blank plates in the fridge and quite a few reasons not to. Obviously you aren't understanding a few things. Why refrigerate blank plates?

Room temp is perfect for blank plates. Actually almost any temp is perfect for blank plates as long as that temp is stable. Fridges fluctuate temp like a mother fucker. Hence the condensation. Just pour a bunch of plates (i do fifty at a time) wrap them in glad or para and store them at room temp until use. Giving the plates a week or more before using them allows any flaws in your procedure to manifest before you throw a potentially clean culture into a potentially dirty plate. We all learn this the hard way. Trust me and you'll save yourself some heart ache.

It is very common to need to do some transfers. It's possible to get clean plates right out the gate but always a good idea to transfer a few times.


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InvisibleTheDoobskerS
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Re: Having troubles with agar [Re: verum subsequentis]
    #26923600 - 09/07/20 06:55 PM (3 years, 5 months ago)

Ah okay. To be honest, idk why I was putting them in the fridge. I was thinking they needed to be in there for whatever reason.

I'm gonna start making more plates at a time and also will keep transferring these plates I have growing.

\thanks


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