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Offlinebuggers
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some questions about agar
    #4584617 - 08/26/05 06:18 PM (11 years, 10 months ago)

i'm planning on getting into agar in order to isolate/ attempt to grow p. caerulipes from a wild print.
i'm thinking about ordering 100 disposable petri dishes and 250 grams of malt extract agar from mycosupply.
i've been told that i need some media antibiotic (available thru sporeworks) in order to prevent bacterial growth on my initial agar plates. whats in this stuff? i've got a bottle of pennicillin - and you've all probably seen the pic with a pennicillin tablet preventing bact. growth on an agar plate. could i crush up 1 or 2 pennicillin pills and put them in when i'm preparing my culture medium?
can the disposable plates be pressure cooked and reused?
just looking for some info and trying to save a few bucks on media antibiotic if i can

thanks


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OfflineIngSocTHC
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Re: some questions about agar [Re: buggers]
    #4584673 - 08/26/05 06:40 PM (11 years, 10 months ago)

With agar if you are growing from mycelium then add some h2o2 to the mix. Sice its in it full grown form it wont kill it but the h2o2 will kill any thing besideds it basicly. So I have read but I dont know from experience.


--------------------
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Offlinebuggers
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Re: some questions about agar [Re: IngSocTHC]
    #4584687 - 08/26/05 06:45 PM (11 years, 10 months ago)

i wont be growing from mycelium as all i have is a spore print. i dont see how h2o2 could fight off bacteria for long periods of time as would be needed for agar plates. the h202 would convert to h20 +o2 rapidly as the agar medium was heated (i think) so it wouldnt do much to stop bacteria present in the spore print from growing on the agar plate


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OfflineAiko Aiko
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Re: some questions about agar [Re: buggers]
    #4584701 - 08/26/05 06:49 PM (11 years, 10 months ago)



--------------------
Man says, "God, show me and I will believe." God says, "Believe and I will show you."


Edited by Aiko Aiko (08/26/05 06:50 PM)


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Offlinebuggers
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Re: some questions about agar [Re: Aiko Aiko]
    #4584722 - 08/26/05 06:56 PM (11 years, 10 months ago)

Quote:

Unfortunately, mushroom spores fall under the category of fungal spores, so they will not germinate on peroxidated agar because they are killed on contact.




Quote:

A good idea would be to order some antibiotic agar from FP to use for germinating spores.




is this what i'm stuck with? what's in the antibiotic agar that makes it antibiotic? could penicillin be mixed in as an antibiotic?


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OfflineAiko Aiko
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Re: some questions about agar [Re: buggers]
    #4584745 - 08/26/05 07:05 PM (11 years, 10 months ago)

Oh yeah! :crazy: If you are comfortrable with your sterile techniques; glove box, flowhood ect... You can probably get away without antibiotic.Another thing you could do is start an LC. It's really easy! Then use the myc. to knock up the agar. :smirk:


--------------------
Man says, "God, show me and I will believe." God says, "Believe and I will show you."


Edited by Aiko Aiko (08/26/05 07:07 PM)


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Offlinebuggers
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Re: some questions about agar [Re: Aiko Aiko]
    #4584762 - 08/26/05 07:14 PM (11 years, 10 months ago)

wouldnt a LC contam just as easily as agar if it was from a Wild print (one that i made from a mushroom that came out of the woods somewhere)

which gives me another idea, however. could i add an antibiotic of some sort to a liquid culture? i've never messed around with agar... but i have done liquid cultures. could i get some clean mycelium by putting spores into a liquid culture medium that was treated with some kind of antibiotic?


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OfflineAiko Aiko
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Re: some questions about agar [Re: buggers]
    #4584773 - 08/26/05 07:19 PM (11 years, 10 months ago)

Start the LC. When it's finished colonizing, you have full blown myc. Then you can knock up a peroxide treated agar dish. Steralizing an LC is very easy. You can either PC it, or just boil PF style. No need for any antibiotic in an LC. GL, Peace! BTW, as long as you made the print in sterile conditions, it should be as good as a cultivated one.


--------------------
Man says, "God, show me and I will believe." God says, "Believe and I will show you."


Edited by Aiko Aiko (08/26/05 07:21 PM)


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Offlinepuwtrip
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Re: some questions about agar [Re: Aiko Aiko]
    #4584794 - 08/26/05 07:28 PM (11 years, 10 months ago)

the problem is, he's using a wild print so it could contam the lc. just lookup an agar tek and get the petri ready. scrape some spores into petri in (glove/flow hood etc). bacterial infection is easy to spot if it's there. once colonized, cut out a teenie bit of uncontammed agar, dunk into h2o2 and drop into sterilized LC.


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Offlinebuggers
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Re: some questions about agar [Re: puwtrip]
    #4584808 - 08/26/05 07:36 PM (11 years, 10 months ago)

my question then is, is there anything other than the "media antibiotic" sold at www.sporeworks.com that i can use in preparation of an antibiotic medium? what is in it?
should i have posted in advanced?


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OfflineAiko Aiko
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Re: some questions about agar [Re: puwtrip]
    #4584987 - 08/26/05 08:35 PM (11 years, 10 months ago)

I could be wrong about this, but if you have an uncontaminated, sporulating mushroom, (wild included) and you take the print in a sterile environment, then you shouldn't have to worry about a contaminated LC.


--------------------
Man says, "God, show me and I will believe." God says, "Believe and I will show you."


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Offlinebuggers
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Re: some questions about agar [Re: Aiko Aiko]
    #4584990 - 08/26/05 08:36 PM (11 years, 10 months ago)

if there were outdoor contaminates on the flesh of the cap, wouldnt they transfer to the spore print?


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OfflineAiko Aiko
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Re: some questions about agar [Re: buggers]
    #4585038 - 08/26/05 08:48 PM (11 years, 10 months ago)

Wouldn't you be able to see them? Look at the contamination forum. There are pics of contaminated mushrooms. It's pretty obvious when a mushroom is infected. If the specimen you took the print from was healthy you should be OK. I will stop there because I have never expireimented w/ a wild specimen. Take a small bit of spores and knock up an LC, and see what happens and let us know.


--------------------
Man says, "God, show me and I will believe." God says, "Believe and I will show you."


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OfflineAiko Aiko
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Re: some questions about agar [Re: Aiko Aiko]
    #4585064 - 08/26/05 08:52 PM (11 years, 10 months ago)

Hey maybe MR. agar can enlighten us. :grin:


--------------------
Man says, "God, show me and I will believe." God says, "Believe and I will show you."


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InvisiblePrisoner#1
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Re: some questions about agar [Re: buggers]
    #4585402 - 08/26/05 10:30 PM (11 years, 10 months ago)

Quote:

buggers said:
i've been told that i need some media antibiotic (available thru sporeworks) in order to prevent bacterial growth on my initial agar plates. whats in this stuff?




generaly it's gentamiacin sulfate, it's not 100% needed, usualy the bacteria
develop because thers too much water on the surface, sometimes condensation
sometimes from culturing with a syringe, to purpose of agar is to isolate
a clean culture from other contaminant like molds and bacteria

a quick tip if your using a syringe, stand the syringe on the plunger and
allow the spores to settle to it for several hours, slowly push out most
of the water leaving a couple of mLs, snce you'll be using agar it's OK to
pull a little air into it to facilitate shaking to disperse the spores,
after your agar plates are cools and ready to use just shake the syringe and
push a couple drops out, then allow a 3rd drop to hang from the tip of th needle
drag that one across the surface of the agar and repeat it with another drop

the purpose is to get some spores on the agars surface while leaving most of the
water behind, cover and seal your plats and you'll be good to go

Quote:

i've got a bottle of pennicillin




use of antibiotic should only be used as a last resort, bacteria can become immune to them, most are not autoclavable and will break down in heat,
I recomend trying 10 plates with and 10 without using 2 seperate batches of agar
in regard to breaking down, they still have a little effectiveness but lose
a great deal of their strength


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Offlinebuggers
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Re: some questions about agar [Re: Prisoner#1]
    #4586793 - 08/27/05 09:25 AM (11 years, 10 months ago)

Quote:

stand the syringe on the plunger and
allow the spores to settle to it for several hours, slowly push out most
of the water leaving a couple of mLs




cool tip! never read that info anywhere good to know. i should get my malt agar, 3 sleeves of dishes, and a bottle of that antibiotic stuff to use on some of my first plates to make sure anything on the print that i dont want doesnt have a chance aganst my mycelium.
thanks.


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