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InvisibleAlkeides
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Registered: 04/11/20
Posts: 61
Agar Transfers not Recovering
    #26670277 - 05/14/20 09:26 AM (3 years, 8 months ago)

Hey fellas I took some MS plates and made ~10 transfers on 5/6 (8 days ago). So far none of the plates are showing any mycelial growth, indicating no recovery. I'm not sure what happened.

Plates use 20g/L BRF agar (same as the MS plate they were drawn from), well water (not flouridated), stored in an incubator at 76F. Following RR instructions I took slices from leading edge of mycelium no larger than a grain of rice. I'm not seeing any contams on either the transfer plates or the control plate I left open in the SAB during the transfer process.

Any ideas what happened?

Second question, the MS plates never showed any rhizomorphic growth only tomentose. The same MS syringes were used to inoculate PF jars some of which are showing beautiful rhizomorphic growth as pictured. Would it be a dumb idea to take a cutting from one of these rhizomorphic jars after birthing, and placing the cutting on an agar plate for expansion and isolation of some rhizomorphic mycelium even if the culture has already gone through anastomosis?



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InvisibleRoger Clemency
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Registered: 03/23/20
Posts: 2,005
Re: Agar Transfers not Recovering [Re: Alkeides]
    #26670426 - 05/14/20 11:09 AM (3 years, 8 months ago)

I’m not sure why they didn’t recover. I’ve never had any problems with that but maybe it had to do with the brf mix. Were the plates you transferred to poured much earlier? Like could they have dried out before you ever used them?

Rhizo or tomentose myc can be manipulated on agar with nutrient levels. That same stuff from the cake would appear differently on the nutrient dense agar


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InvisibleAlkeides
Child

Registered: 04/11/20
Posts: 61
Re: Agar Transfers not Recovering [Re: Roger Clemency]
    #26670476 - 05/14/20 11:46 AM (3 years, 8 months ago)

No I think I poured those plates only a day or two before and they were kept refrigerated inside a tupperware deli container inside a gallon ziploc bag. I fit about 40 in there cus they're only 35mm.

I should also mention each one is wrapped twice with cling wrap, maybe that's reduced GE to the point of stalling but with such a small sample shouldnt there be at least some visible activity just off the gas that was trapped inside the volume of the plate?

I have some rice soak water leftover from vtek i could try to pour some plates with, but if the agar recipe was the factor here I wouldn't have gotten any growth off the orginal MS plates which have now exploded....


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InvisibleAlkeides
Child

Registered: 04/11/20
Posts: 61
Re: Agar Transfers not Recovering [Re: Alkeides]
    #26673114 - 05/15/20 05:14 PM (3 years, 8 months ago)

Hate to bump but I'm a little stumped here. Still no recovery.

In addition I have some other old spores that germinated on filthy plates and while two of those transfers have recovered they both have also stalled...


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