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OfflineScarg
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Mycellium not jumping on to new agar
    #19045939 - 10/28/13 12:41 PM (10 years, 3 months ago)

I work in front of a flowhood.  My agar recipe is 10g agar, 15g light malt extract, and 1-2g brewers yeast per 500ml of water.  Never had a problem with agar before. 

In the last month ive started about 60 petris from spores (some PE, some another cube variety).  All of those dishes took off and I started transfering the thickest sectors from each dish...lots and lots of dishes). 

Some of those transfers took off again, and some did not.  I transferred the ones that did again, and so far all of them have not taken off.

All of my cultures now (both 1st and 2nd transfer) will not jump on to the new agar plates. I even tried making a new batch and putting the colonized wedges that dont want to jump over, on to the new plates (thinking that maybe I forgot an ingredient or something on the previous batch).  Still nothing.

Has anybody ever encountered this?  Why is this happening?  Is there a fix?


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InvisibleSpitballJedi
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Re: Mycellium not jumping on to new agar [Re: Scarg]
    #19045991 - 10/28/13 12:55 PM (10 years, 3 months ago)

15g LME seems rich to me. Try using only 10g and ditching the yeast.


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OfflineScarg
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Re: Mycellium not jumping on to new agar [Re: SpitballJedi]
    #19059669 - 10/30/13 07:57 PM (10 years, 2 months ago)

Ive been doing some research and have read that LME has a shelf life.  Has anybody ever heard of this?  Maybe the year and a half old bag Ive got just isnt viable anymore for mycology purposes?

Im going to buy a fresh bag and see what happens, but Im still interested to hear everyones experiences.  It just doesnt make sense that it worked and worked and worked...then all of a sudden it stopped.


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InvisibleMUSH HEAD420
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Re: Mycellium not jumping on to new agar [Re: Scarg]
    #19060188 - 10/30/13 09:04 PM (10 years, 2 months ago)

I would say it is your culture that is not viable, though it could be the recipe, or it could be you screwed up the recipe.

Sometimes the mycelium just looses the will to live after so many transfers  :shrug:

I would guess that if after the first transfer you notice good characteristics that would be the point where you want to use it.


Edited by MUSH HEAD420 (10/30/13 09:08 PM)


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OfflineRogerRabbitM
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Re: Mycellium not jumping on to new agar [Re: Scarg]
    #19060419 - 10/30/13 09:28 PM (10 years, 2 months ago)

That recipe is too rich, so cut it back.  Another thing to watch for is if your water supply changed pH.
RR


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OfflineScarg
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Re: Mycellium not jumping on to new agar [Re: RogerRabbit]
    #19060483 - 10/30/13 09:36 PM (10 years, 2 months ago)

Could be the culture, but seems unlikely.  The mycellium is only about a month old from spores, and its all of my cultures from a couple of different varieties.  It all just stopped one day...I also bought a new brand of agar around the same time that I noticed this started happening. That also seems like an unlikely explanation, but it is pretty coincidental.  Ill also try buying my old brand of agar along with new LME.  Hopefully itll work.  Im trying to get some good isolates, and be done with MS for good.

Quote:

RogerRabbit said:
That recipe is too rich, so cut it back.  Another thing to watch for is if your water supply changed pH.
RR




I had suspected this before and made a couple of new batches with just 10g LME, and no yeast.  So far it hasnt helped.  Whats an ideal PH?


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OfflineRogerRabbitM
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Re: Mycellium not jumping on to new agar [Re: Scarg]
    #19060519 - 10/30/13 09:41 PM (10 years, 2 months ago)

5.5 to 6.
RR


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InvisibleMUSH HEAD420
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Re: Mycellium not jumping on to new agar [Re: Scarg]
    #19060522 - 10/30/13 09:41 PM (10 years, 2 months ago)

that sounds low, I thought the ph should be at least 6.5

Edit: I see now that above 6.5 ph is reserved for casing.


Edited by MUSH HEAD420 (10/30/13 09:45 PM)


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