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Offlinepsycho_nauticus
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Agar wedge failed to inoculate and recover...?
    #26937647 - 09/15/20 05:37 PM (3 years, 4 months ago)

Hey guys, I inoculated a couple jars with some agar wedges, had clean cultures. This is the first time I have experienced this, but they have failed to inoculate my jars and fuzz up and spread. Some mycelium came off whenever I had to jiggle them somewhat to get the agar to the middle and bottom, that's happened before, but this time they never recovered and fuzzed back up and failed to inoculate my jars... Why would this happen and how is this possible?

P.S. Does it have anything to do with PC'ing my grains at 16 psi? The only reason I say this is because my 15 lb weight doesn't start shaking until it hits 16 psi, so I think the gauge may be off, but I am going by the weight instead of the dial, but lets just say for the sake of argument that I PC'd them at 16 psi, will that affect the inoculation of the agar?


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Offlinesmalltalk_canceled
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Re: Agar wedge failed to inoculate and recover...? [Re: psycho_nauticus]
    #26937715 - 09/15/20 06:07 PM (3 years, 4 months ago)

Could be anything really. We'd need pictures of the plates you used, and info about what you did in the SAB. Sounds like your spawn was far from healthy if there was no leapoff on grains. When transfering healthy agar, you will have growth within 3 days on grains, usually before that too.

Could be your spawn was bacterial. Bacterial spawn to grain uses a lot more time before it takes off, and the bacteria probably follows.

If you place a very small agar piece on top of grains, it can dry out sometimes too.


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OfflineOne of Us
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Re: Agar wedge failed to inoculate and recover...? [Re: psycho_nauticus]
    #26937717 - 09/15/20 06:08 PM (3 years, 4 months ago)

How long has it been? Was the plate stored in the refrigerator before inoculation? Outside of that, I'm not sure what would cause that.

Maybe there was still a lot of bacteria that survived somehow (new bag of oats with higher bacterial load, or improper venting?), and the mycelium really didn't want to leap off into that.

:shrug:


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Offlinepsycho_nauticus
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Re: Agar wedge failed to inoculate and recover...? [Re: One of Us]
    #26937789 - 09/15/20 06:39 PM (3 years, 4 months ago)

I always vent my PC for 10 minutes before applying the weight and starting the cycle. I prep my rye like I always do... I believe my grain must be good because I inoculated another jar I PC'd in the same cycle all together with another Cubensis variety, and it's doing fine... I'm literally trying to find out an explanation for this... I would understand if it was a straight MS spore syringe to grain, but doing agar work and using a T3 clean transfer, I don't understand... I did not take pics of the plate :frown:

Could it be from PC'ing grain at 16 psi instead of 15 psi? Maybe it makes a difference in Cubensis varieties?


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Edited by psycho_nauticus (09/15/20 06:57 PM)


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