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OfflineThatDaltonGuy
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Registered: 04/07/04
Posts: 213
Loc: USA
Last seen: 17 years, 6 months
MEA CONTAM
    #2543934 - 04/09/04 04:50 PM (17 years, 7 months ago)

ok this has never happend to me before...

about a month ago i made some MEA in my PC and sealed it in a bunch of plastic viles... when came to pour my dishes, i scraped the stuff into a double boiled with a sterile knife and boiled the mea once again for about 30 minutes... then poured into 6 dishes and set in my closet closed off for about 3 hours to dry... in the mean while i sprayed my entire closet down with h202 and isopropyl mix and was wearing a dust/fume mask... i had some portobello in the closet in a plastic bag that had been decomtaminated by washing with h202 and then rinsing off in boiling water.... these were store bought portobellos and possibly could have had contams on them... but ive never had any problems with these methods before... i then proceeded to cut little pieces of the gills off and place on the agar... once again with sterile knife and sterile conditions... well its been about 4 days, and everything has been looking good until today, where patches of green mold have bursted out all over the place... does anyone have any links to any better methods of germinating on MEA? should i just make syringes instead of germinating on MEA? feedback please!


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OfflineMAIA
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Registered: 04/27/01
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Re: MEA CONTAM [Re: ThatDaltonGuy]
    #2550384 - 04/12/04 06:35 AM (17 years, 7 months ago)

Quote:

i had some portobello in the closet in a plastic bag that had been decomtaminated by washing with h202 and then rinsing off in boiling water .... these were store bought portobellos and possibly could have had contams on them...




This is not a safe method if you consider the mushrooms were contaminated. Working with agar is a very delicate thing, if you put any contaminated material, then most probably you'll end up with a contaminated dish. You can try saving those with some antibiotic or germicide but if you don't have any experience, it will be very difficult.
Also, do you use a glove box ? It's important you have one if you're going to work with agar.

Quote:

should i just make syringes instead of germinating on MEA?




You should give it a try.

Just to add that it's very important when you select and collect the mushrooms sample to clone in the agar dish. It seems you usually use the gills but you can also cut small pieces from the stem and clean it with some h2o2. Forget about rinsing the sample in boiling water, it does more harm than good.

MAIA


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Spiritual being, living a human experience ... The Shroomery Mandala



Use, do not abuse; neither abstinence nor excess ever renders man happy.
Voltaire


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