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Amazon Shop for: ½ Pint Jars, Agar, Petri Dish

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Offlinexburn
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Agar to LC
    #4203357 - 05/21/05 02:25 PM (11 years, 6 months ago)

THis is a two parter.

When going from agar to lc how large a piece do you use?

and when doing a syringe to agar, if you put to much water form the syringe and it turns milky after three days. the water on the plate that is. Is that just the monycrons and dikyrons?


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OfflineRogerRabbitM
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Re: Agar to LC [Re: xburn]
    #4203515 - 05/21/05 03:01 PM (11 years, 6 months ago)

I've used a whole petri dish to inoculate one liter of liquid culture solution.

I never use more than one drop from a spore syringe right in the very center of the petri dish. You don't want a huge amount of spores germinating. That defeats the purpose and makes strain isolation that much harder. The milky water might be germinated spores or it could be bacteria. Keep a close watch.
RR


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Offlinexburn
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Re: Agar to LC [Re: RogerRabbit]
    #4203571 - 05/21/05 03:20 PM (11 years, 6 months ago)

thanks


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OfflineBlue Helix
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Re: Agar to LC [Re: xburn]
    #4216269 - 05/24/05 11:00 PM (11 years, 6 months ago)

I agree with RogerRabbit. Spore solutions on agar work best at about 100 um (1/10th a CC) in the center of the plate.

I use half pint jars with a hole in the metal lid and covered by a filter patch. I pressure cook them with the agar inside and then sit the jars on the shelf until the condensation goes away (about 3 days).

Once the condensation has gone away, I inoculate the plate. I draw out the solution from my vacutainer storage test tube with a 1/2ml syringe sporting a 29-gauge needle. I clean the room air with Oust, allow the Oust to settle, and then remove the filter patch exposing a small hole to inject the solution. I inject 100 um (average drop) and put back on the filter patch with the ring. Then I sit the jar out in the open (not in an incubator) for a day or two until the drop completely evaporates. I then incubate the jar inverted at 86F inside a bucket-in-bucket setup. I find that the jar inversion prevents the agar from sweating and causing condensation and also promotes a more fluffy mycelium growth.

Remember this: standing water is your enemy in this hobby. It doesn't matter how sterile you think your grain is or your agar plate, if you see standing water, you will eventually have bacterial problems. That is why it is critical that the condensation does not form and the spore solution evaporates.


Edited by Blue Helix (05/28/05 03:00 AM)


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Re: Agar to LC [Re: Blue Helix]
    #4217083 - 05/25/05 03:10 AM (11 years, 6 months ago)

Very nice BH, sounds like a great way to do agar... The agar never falls since its inverted? I assume its to light..


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Re: Agar to LC [Re: scatmanrav]
    #4217170 - 05/25/05 03:39 AM (11 years, 6 months ago)

I have never had the agar fall and I've been incubating agar jars inverted for several years. I think it's because the agar is so sticky. I have seen the agar shrink from the sides when it dries out, but since the jars are sitting on their own filter patch when inverted, they won't dry out in that orientation.


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Re: Agar to LC [Re: scatmanrav]
    #4217178 - 05/25/05 03:44 AM (11 years, 6 months ago)

And I wanted to add that you can also, of course, store mycelium in sterile water in a vacutainer too for years in the refrigerator. That can be great if you clone some mycelium from a champion shroom into a live culture. You'll get perfectly consistent results without taking all that time to isolate (although isolation isn't really necessary anyway with such a robust mushroom like cubes).


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Amazon Shop for: ½ Pint Jars, Agar, Petri Dish

Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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