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Invisiblesci33
AmbitiousScientist
Registered: 11/11/03
Posts: 186
Strength of Kanamycin?
    #2814552 - 06/21/04 05:43 PM (12 years, 5 months ago)

Friend of a friend got me some kanamycin (antibiotic) from a lab. Been mixing it into my agar for a while with great results for multispore cubensis germinations. Mixed it into some PDYA a while ago and spread some Pan cyan on it - nothing germinated. Is it possible its very strong and weak myceliums won't grow on it? Tampanensis seems to be doing fine on it though.


--------------------
Looking for:
Ps. weilii, Ps. samuiensis, Ps. hispanica, Pan. cyanescens (any strain I don't have)

Have:
Ps. cubensis, Ps. tampanensis, Pan. cyanescens (Australian, Tai-KS, Hawaiian, Jamaican, Ban Lipa Noi)

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OfflinePsilygirl
cyan goddess
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Re: Strength of Kanamycin? [Re: sci33]
    #2814793 - 06/21/04 07:28 PM (12 years, 5 months ago)

can you eliminate any other reason that it didn't germinate?  if it worked well for other strains, even a weak one such as Tampanensis, then it seems that it should work fine.

explain the time frame, how many plates you did, etc... was it a separate batch of PDYA that you made for the Pan cyan? 

how much of the kanamycin do you use?

i'll try and help if i can

:sun:
Psily


--------------------
"Love says 'I am everything.' Wisdom says 'I am nothing.' Between the two, my life flows."


Puget Sound Mycological Society


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Offlinerootkey
Canada Rules
Registered: 04/18/02
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Re: Strength of Kanamycin? [Re: sci33]
    #2814885 - 06/21/04 07:54 PM (12 years, 5 months ago)

I don't know much about the uses of Kanamyacin in regards to mushroom growing but i know it is a popular Fish Medication for anyone who has serious interest in getting your hands on some. It is used in the fishkeeping hobby to treat a disease called Columnaris along with many other bacterial diseases. Any serious fish/petstore should have it in stock, or be able to get thier hands on it. (it looks like cocaine) LOL.


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Invisiblesci33
AmbitiousScientist
Registered: 11/11/03
Posts: 186
Re: Strength of Kanamycin? [Re: rootkey]
    #2814921 - 06/21/04 08:08 PM (12 years, 5 months ago)

I did about 20 plates, on a few occasions, but that one was from a batch of 20. I unfortunately had to eye out a knifetip of the kanamycin so I don't know the measurement.. still waiting on getting a milligram scale.


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OfflineSuntzu
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Re: Strength of Kanamycin? [Re: sci33]
    #2814944 - 06/21/04 08:14 PM (12 years, 5 months ago)

Was the panaeolus that you spread in the form of spores or mycelium? How old was it in any case? Are you sure that the culture/spores are currently viable? Pans are the quickest to die out, even a 'nice' looking plate that's been around awhile may not be viable.
Kanamycin should not affect Pan growth differentially, though at extreme concentrations anything can happen. I suppose it's possible that the mitochondria of Pan. mycelium is susceptible to kanamycin [mitochondria being the bacterial slaves of us eukaryotic bad-asses].
Before we can be really suspicious, the spores/culture would have to be shown viable on a non-kanamycin media.
As Psilygirl asked, what concentrations were you using?

*edit* You beat me to it!


Edited by Suntzu (06/21/04 08:15 PM)


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Invisiblesci33
AmbitiousScientist
Registered: 11/11/03
Posts: 186
Re: Strength of Kanamycin? [Re: Suntzu]
    #2814964 - 06/21/04 08:19 PM (12 years, 5 months ago)

This was a hawaiian pans print. It never had a great germination ratio, but it definitely did germinate before. I think I will just do up a few more plates before jumping to conclusions.


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OfflinePsilygirl
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Re: Strength of Kanamycin? [Re: sci33]
    #2815113 - 06/21/04 09:08 PM (12 years, 5 months ago)

sounds like a good idea :thumbup:

maybe do some plates with kanamycin, some without--keeping all other factors the same.

:sun:


--------------------
"Love says 'I am everything.' Wisdom says 'I am nothing.' Between the two, my life flows."


Puget Sound Mycological Society


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OfflineSuntzu
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Re: Strength of Kanamycin? [Re: Psilygirl]
    #2815166 - 06/21/04 09:42 PM (12 years, 5 months ago)

Or if you have the plates to spare, maybe a few different dilutions of kanamycin. This could easily be done with two bottles/flasks of agar. Add kanamycin to one as you did before, pour a plate. Mix varying amounts of the kanamycin and non-kanamycin agar for subsequent plates. This could really prove that it is inhibitory [and at approximately what concentration].
Might be a pain in the ass, depending on the tools at hand.


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InvisibleAlkaloids
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Registered: 11/16/98
Posts: 743
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Re: Strength of Kanamycin? [Re: sci33]
    #2836771 - 06/28/04 04:56 PM (12 years, 5 months ago)

I would recommend using the kanamycin at concentrations of 50-100 mg/L. The temperatures you are using for incubation and the total length of the incubation period will determine, to an extent, what concentrations you will want to use, but 50mg/L is a good amount and I don't imagine you would ever need to go above 100mg/L as that won't really provide any selective advantage.


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Offlinetubervegan
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Registered: 07/02/04
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Last seen: 2 years, 6 months
Re: Strength of Kanamycin? [Re: sci33]
    #2852020 - 07/02/04 05:53 PM (12 years, 5 months ago)

Cubensis is a very un-hardy mushroom, mold could probably deal with the kanamycin more easily than cubensis could. The reason why mold and our usual contams grow everywhere is because they are strong, cubensis grows in few places because it is a weak species so if you are using fungicides basically if you can get cubensis on it then you can grow mold on it and all you are going to end up with are contaminated cultures that would produce really shitty msuchrooms anyways because they are going to have to deal with the fungicides the entire time they are growing.


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