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InvisibleTHEDANGLER
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antibiotic agar
    #4034650 - 04/09/05 04:48 PM (12 years, 5 months ago)

i figured this would be a good place for this question...

i made up some dogfood/oatmeal/dextrose/peptone agar and a bunch of my little canning jars i use for petries filled with water so im going to use the ones that didn't but that will leave me with a big quart jar of antibiotic agar, the atibiotic im useing is amp or kan which ever i choose, will it be ok if i keep the big jar in the freeze and just melt it down later to use? will the antibiotics survive the melt down?


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Offlinemattymonkey
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Re: antibiotic agar [Re: THEDANGLER]
    #4036435 - 04/10/05 08:28 AM (12 years, 5 months ago)

don't you normally put the antibiotics into the autoclave? im sure they would survive remeltdown.. what would you use, a hotplate?


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Invisibleohmatic
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Re: antibiotic agar [Re: mattymonkey]
    #4036541 - 04/10/05 09:54 AM (12 years, 5 months ago)

i have written a post about the use of antibiotics in agar that you should mind.
i dug it up, here you go:
Quote:

jeppa thats it, autoklaver her at 18psi (or greater) for an hour and whats there shall be dead.

anyways, conecerning this fact, guides like http://www.kitchenculturekit.com act incredibly
irresponsible by stating:
Quote:


Allow media to cool until mixture is warm to touch, however, not solidified. Add 0.5 cc of Gentamicin Sulfate per 10 cc of solution (1.5 cc per cup)



(source http://www.kitchenculturekit.com/Mushroom/agarplates.htm)

especially since this guide aims at newbies, it will have already
caused a lot of them to breen mad contams flying around out there for sure.

as stated above, use antibiotics only in EMERGENCIES and never when its
not absolutely needed.
peace ohm :mushroom2:





concerning your question, i am quite sure that the re-heating will destroy the
antibiotic even though tehre are certain kinds (imho also gentacimin) that
can actually withstand autoklaving.

MIND my post above, if you breed a resistant contam strain your going to get
majorely buttfucked.
peace ohm :mushroom2:


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OfflineRogerRabbitM
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Re: antibiotic agar [Re: ohmatic]
    #4036587 - 04/10/05 10:34 AM (12 years, 5 months ago)

Also bear in mind, the antibiotics will only help to protect against bacteria. They will be of no use against clandosporium, aspergillus, penicillium, trichoderma, etc., etc.

As shown above in ohm's post, allow the agar to cool before pouring. If water is condensing in your dishes, it sounds as if you poured the agar while it was still too hot.
RR


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Invisibleohmatic
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Re: antibiotic agar [Re: RogerRabbit]
    #4036594 - 04/10/05 10:40 AM (12 years, 5 months ago)

Quote:

RogerRabbit said:
Also bear in mind, the antibiotics will only help to protect against bacteria.  They will be of no use against clandosporium, aspergillus, penicillium, trichoderma, etc., etc




jeah major factor.
you maybe want to quit messing totally with antibiotics and look into
using peroxided agar instead.

heres a link, check it out: http://www.shroomery.org/index/par/23462

btw, cool to read more of your posts around here recently rodger :smile:
peace ohm :mushroom2:

*edit*
if you plan on raising culture from spores, youll have to 'pre-breed' them
on non-peroxided agar b/c peroxide will roast them spores instantly.
after you got one motherculture going, you can then sector and transfer to the peroxided dishes.


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Edited by ohmatic (04/10/05 10:42 AM)


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InvisibleTHEDANGLER
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Re: antibiotic agar [Re: ohmatic]
    #4037454 - 04/10/05 04:51 PM (12 years, 5 months ago)

thanks for the help i guess im use antibiotics to clean up some cultures i used about 1yr ago...last time i tryed peroxide i think it brought some contams in with it..what temp does amp and kan( i dont rember their full name) break down at and what temp does agar melt at?


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OfflineAeolus1369
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Re: antibiotic agar [Re: THEDANGLER]
    #4037497 - 04/10/05 05:01 PM (12 years, 5 months ago)

I was always told to add kanamycin to agar at no hotter than 50C (umm, about 120F I think) and agar melts at well above that. I doubt all of the antiobiotic would break down if it was cooled in a reasonable amount of time though.


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InvisibleTHEDANGLER
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Re: antibiotic agar [Re: Aeolus1369]
    #4037514 - 04/10/05 05:04 PM (12 years, 5 months ago)

yah but im saying the agar has already cooled so should i just melt the agar or pc it again befor i use it?


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OfflineAeolus1369
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Re: antibiotic agar [Re: THEDANGLER]
    #4037554 - 04/10/05 05:12 PM (12 years, 5 months ago)

Oh sorry I misunderstood. If you want to add antibiotics to already prepared plates, you can just dissolve the antibiotic in sterile water and spread that out on the plate. It will be absorbed and the antibiotic will diffuse throughout the agar pretty quickly.

Alternatively, you can melt down a small amount of agar and mix the antibiotic in there and then pour that on top of the plates.

Does that help?


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Invisibleohmatic
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Re: antibiotic agar [Re: Aeolus1369]
    #4037631 - 04/10/05 05:34 PM (12 years, 5 months ago)

Quote:

Aeolus1369 said:
Alternatively, you can melt down a small amount of agar and mix the antibiotic in there and then pour that on top of the plates.





i instantly had to think of the 'agar sandwich' tek when i read that.
still, it isnt clear for me why the threadstarter even wanted to use
any antibiotic in the first place ...
peace ohm :mushroom2:


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InvisibleTHEDANGLER
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Re: antibiotic agar [Re: ohmatic]
    #4038110 - 04/10/05 07:58 PM (12 years, 5 months ago)

because i have them and wtf else am i going to use the antibiotics for...yes the agar sandwich thing is a very good tek but im not trying to do all that...the antibiotics were already put into the masterjar of agar and 4 plates were pour...the agar hardened and its in the freezer now...should i re-pc the jar or will it be fine to just melt?


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Invisibleohmatic
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Re: antibiotic agar [Re: THEDANGLER]
    #4039563 - 04/11/05 02:19 AM (12 years, 5 months ago)

Quote:

THEDANGLER said:
because i have them and wtf else am i going to use the antibiotics for...yes the agar sandwich thing is a very good tek but im not trying to do all that...the antibiotics were already put into the masterjar of agar and 4 plates were pour...the agar hardened and its in the freezer now...should i re-pc the jar or will it be fine to just melt?




if you poured without a flowhood - pc it again.
im quite sure that the antibiotic will get destroyed in the pc.

also, you dont realise what can happen if you breed a resistant strain,
youll definitely come to think of my advice once you got some mold flying
around youve never ever seen ...
peace ohm :mushroom2:


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InvisibleTHEDANGLER
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Re: antibiotic agar [Re: ohmatic]
    #4040171 - 04/11/05 09:59 AM (12 years, 5 months ago)

yah well what if your peroxide perduces a resistant strain...ever think of that? the plates were poured in a glove box..


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Re: antibiotic agar [Re: THEDANGLER]
    #4040182 - 04/11/05 10:07 AM (12 years, 5 months ago)

Quote:

THEDANGLER said:
yah well what if your peroxide perduces a resistant strain...ever think of that? the plates were poured in a glove box..




its impossible as spores are instantly grilled.
glvoeboxes are semi sterile environments,
you can be good but tehre is no 100% guarantee.
peace ohm :mushroom2:


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InvisibleTHEDANGLER
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Re: antibiotic agar [Re: ohmatic]
    #4040216 - 04/11/05 10:24 AM (12 years, 5 months ago)

yeah but what if their is a couple spores that arnt destroyed...aka they evolve....nothing is impossible..


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Offlinescatmanrav
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Re: antibiotic agar [Re: THEDANGLER]
    #4040451 - 04/11/05 12:00 PM (12 years, 5 months ago)

I disagree, many things are impossible. Take a physics class :smile:


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Re: antibiotic agar [Re: scatmanrav]
    #4040951 - 04/11/05 02:10 PM (12 years, 5 months ago)

Quote:

scatmanrav said:
I disagree, many things are impossible. Take a physics class :smile:




:wink:

let the water flow upwards god damnit !!!
peace ohm :mushroom2:


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Re: antibiotic agar [Re: ohmatic]
    #4041578 - 04/11/05 05:22 PM (12 years, 5 months ago)

their are spores out their that can survive pressure cooking im sure theirs a couple species that can survive h202


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Offlinemattymonkey
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Re: antibiotic agar [Re: THEDANGLER]
    #4042591 - 04/11/05 09:23 PM (12 years, 5 months ago)

anyway, about this antibiotic thing.. i have this hericium im trying to start from spore.. i think its hericium colorodies(sp?) i gathered the spores last fall when i didnt have a lab setup, and started working with them just a few weeks ago.. they germinated but were very slow, super slow compared to the oyser spores i started from...
i have this weird yellowish tint now on the plates, it seems to have come with the myc.. imtrying to isolate out any strain i can as i don't have the spore print anymore.. if i have some myc growing on a weird dish, can i use AB to transfer it to a different dish, and just propogate the myc? would this h2o2 thing work better?
thanks


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Re: antibiotic agar [Re: mattymonkey]
    #4042871 - 04/11/05 10:29 PM (12 years, 5 months ago)

i think it works about the same just breaks at a higher temp


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