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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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Agar Medium
    #534722 -

I have just started growing on a home made PDA medium.
I didnt properly filter the Potatoe and dextor broth so there is a very small trace of potatoe matter in there and i did not weight the potatoes but the other ingredients are at the correct weight. I did forget to use distilled water but because of the pressure cookin of the medium wouldnt that evaporate any chlorine in the water.

It has been about 7 days since i have inoculated it with Psilocybe cubensis "Ecuador" from mush mush.

So far i have seen no contamination or any other growth except on one dish which looks like it has starch stains on it. Also there is a small amout of condensation on the petri dishes. So now for the questions.

How long does it take for spores to grow in to mycelium ?

Should the dishes still be wraped after the inoculation of the spores ?

Does the condenstaion or extra liquid caused by the syringe effect growth rate ?

I have also have about half of litre of medium left in a sterile bottle how long will this last for ?

Thanks


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Fighting the man the best way I can.

Edited by jezu (01/29/02 06:02 AM)

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Re: Agar Medium [Re: EvilGir]
    #534899 -

Lo
First don't worry about your PDA formula.
I've started agar 2 weeks ago, using a potato/carrot/yeasts agar without weighting anything (exept agar-agar).
First batch lost to black pin mold in 2-5 days
Second batch innoculated in glovebox 5 days ago with a print=> you can see tiny spots of tiny ropy things...

>Should the dishes still be wraped after the inoculation of the >spores ?
Wrapping the dishes depends on where u store them. If in clean filtered box, don't need to wrap.
I now store them unwrapped in my glovebox (which is big enough ...) and had no contams pb so far.
The "witness" dish (one which is not innoculated) is clean!!
But wrapping is a precaution.

>How long does it take for spores to grow in to mycelium ?
I consider 10 days to be the limit but if no contam on dish, wait and see.

>Does the condenstaion or extra liquid caused by the syringe >effect growth rate ?
Mycelium can be hurt by water in dishes but just wipe it out.
If you innoc with a syringe, 2 to 10 dropplets should be enough so the extra-water dries quick.

>I have also have about half of litre of medium left in a sterile bottle how long will this last for ?
Depends where and how you store it.....resterilize before use...

Hope my tniy experience helped you a bit :smile:

++
Duss69 


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Support the FSRE

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Re: Agar Medium [Re: duss69]
    #535647 -

Last night i have removed all the wate from the petri dishes under aseptic conditions. Next i found a plant incubator clean it with bleach and then
placed the petri dishes inside it.

In the morning the dishes had seemed to of gained more water say about 0.3ml.
And looked way to wet for anything to grow. Even though when i inoculated them I used 1 small drop.

I have noticed that the only dish that has remained dry has been contamanated.
So i hve decided to throw the 5 dishes out and start from scratch.

So Does anyone have any suggestion on how to avoid water contam.
As this seems to be the main problem. I also think that I am makeing my nutrient wrong so anyone have any suggestion and tips for pouring and mixing nutrient.

Thanks


--------------------
Fighting the man the best way I can.

Edited by jezu (01/30/02 02:39 PM)

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Re: Agar Medium [Re: EvilGir]
    #536654 -

By the wya, your agar. The stuff to be used, i a/c in a beaker, the stuff to not be used, is a/c in a mason, then when it comes out of the a/c, the lid is screwed down, and not taken off until its used again (reheated on stovetop)
anyway... Glass pitres tend to condensate more, because they hold heat better, and causes them to condensate.
make sure you store them upside down when not in use..
and make sure they are (when in use) in an area that is not changing in temperature.
i made this mistake too.
i had them in the fridge, then brought them out to the room for only 20 mins, and then inoculated, then put them in the incubator
they condensated pretty bad. so basically, let em warm up in your incubator for awhile, inoculate, then put them right back in.
btw, try to use plastic pitres, instead of glasses or whatever you may use.

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Re: Agar Medium [Re: cookiewhore]
    #537232 -

quick ?
if i use LB, should i add anit-b
also how wet should LB be?


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Yo Quiero FSR !

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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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