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 Richard_D_James
enthusiast
Registered: 04/27/01
Posts: 242
Last seen: 16 years, 7 months
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 MEA - Help! 
#320079 - 05/18/01 11:16 AM (23 years, 11 months ago) |
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I recently poured my first agar plates. I scraped B+ into 3 of the plates. Now 7 days later, after incubating @ 81 F, the only thing that exists is extremely cottony mycelium. It is only a very thin layer of mycelium, not rhizomorphic at all. The agar media has begun to pull away from the sides of the petri dish. I haven't seen any new signs of growth for the past 3 or 4 days. Also, the top of the dish has been condensating like a fat girl in the kentucky sun.
any ideas?
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 dimitri211
Carpal Tunnel
Registered: 04/27/01
Posts: 2,248
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this belongs in the cultivation section there is nothing advanced about agar----but relax your are trying to rush---I will help you out most of the time when you 1st put spores on agar it willl usually be cottony----Because the agar is pulling away from the sides you need to do a transfer NOW -your moisture is evapoating and that will lead to the unsitely death of your mycelium---to avoid this problem in the future you can buy parafilm (fungi.com) or I use a 3m brand of eletrical tape and run it around the petri dish GOOD LUCK
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gray1
addict
Registered: 04/30/01
Posts: 430
Loc: brooklyn
Last seen: 23 years, 16 days
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what were you're other incubation conditions?
it seems odd that you were getting condensation and dehydration of the agar simultaneously as you described. how was the agar solution made/poured?
you might want to try with lids off in a sterile incubator type environment with sufficient humidity.
really though, it takes a lot of dehydration for the agar to dry out and pull away from the plates, my preliminary thoughts are that therein lies the main problem
post more specific details and i'll try to analyze some more
c12h16n24ohdmt
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Triple_3
member
Registered: 03/21/01
Posts: 99
Last seen: 23 years, 9 months
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Re: MEA - Help! [Re: gray1]
#320148 - 05/18/01 01:31 PM (23 years, 11 months ago) |
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adding ground grain of substrate you are using, i.e. rye grain
helps to promote rizomorphs and helps get a head start for myco's developing enzymes for the particular substrate you are using.
deeper pouring helps it not to evaporate.
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egghead
veteran
Registered: 04/25/01
Posts: 1,054
Loc: Milky Way
Last seen: 23 years, 1 month
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Rich.. Watching mycelia grow on agar is probably my fav part of the hobby. Do you have a microscope?
If you're having trouble w/ your media drying out and/or don't want to fool w/ tape, etc.. try using the short fat wide-mouth 1/2 pint jars. The idea for this came from a friend who is missing named Karl Brooks. To make slants, I put'em back in their box and then tilt the box while they cool. Some of the nice things about them are:
- pre-pour plates before sterilizing
- control moisture by loosening lid (I like the plastic ones)
- easier to handle
Another way of combating the condensation is to flip the petri over and let the mycelia grow hanging upside-down.
* Contributor to the Free Spore Ring
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Where there's skill, there's a better way..
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Richard_D_James
enthusiast
Registered: 04/27/01
Posts: 242
Last seen: 16 years, 7 months
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I ordered plastic petris from edmund scientific, 100x15mm, washed them out with alcohol b4 i used them. I took 21 grams of MEA from fungi.com, mixed with 500mg of water, stirred well, and Pressure cooked for 35 mins @ 15 psi. Poured plates in my glove box. They were still pretty warm when I poured them.
This IS advanced compared to what shit gets posted in the cultivation forum. ""How many B+'s should I eat before I get fucked up?" blah blah.
About using 1/2 pint's, how should I enable gas exchange? Pafafilm or what not?
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egghead
veteran
Registered: 04/25/01
Posts: 1,054
Loc: Milky Way
Last seen: 23 years, 1 month
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Hmm.. 5 grams of organic flour (preferrably from the grain you'll transfer to) probably wouldn't hurt that recipe.
If you want to exchange gas (I don't normally find it necessary), just loosen the lids to let moisture escape. Not exactly state of the art, but should get you by.
* Contributor to the Free Spore Ring
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Where there's skill, there's a better way..
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