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Offlinepluteus
level-9 deviant

Registered: 08/12/03
Posts: 170
Loc: London area, UK
Last seen: 15 years, 2 months
Re: Agar recipe for cloning oysters [Re: G a n j a]
    #2172061 - 12/12/03 03:38 AM (18 years, 1 month ago)

I use 30% and fried my cultures to death first time round...
that's 0.6 ml / l for 30% folks

for cloning from wild, I prepare antibiotic media, peroxidated media, and also antibiotic + peroxidated media. I have the most success with the antibiotic stuff, but every so often I find a species that's easier to isolate on combination media. e.g. bird's nest fungi

Antibiotic + peroxidated media also seems to work quite well for cloning from rhizomorphs, such as you find growing in woodchip beds underneath colonies of P. cyanescens. Sometimes I quickly rinse the rhizomorph in dilute ethanol first.


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InvisibleG a n j a
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Registered: 12/03/02
Posts: 7,860
Loc: Zone ate
Re: Agar recipe for cloning oysters [Re: pluteus]
    #2172107 - 12/12/03 04:05 AM (18 years, 1 month ago)

My 17% goes in at .84ml per every 500ml.
Do you know of any antibiotic addatives I
can get hold of in the uk ? As that would
be real handy for strating spores from
wild picked mushrooms :cool:

Do antibiotic's stop bacteria ?


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Offlineragadinks
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Registered: 10/20/03
Posts: 1,297
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Re: Agar recipe for cloning oysters [Re: G a n j a]
    #2173352 - 12/12/03 06:45 PM (18 years, 1 month ago)

I do not know what the situation is like in the UK. I got gentamycin from my local doc ( but he's a relative ... :smirk: ). You might also be able to get it from a vet. I have heard that you can get antibiotics in some pet shops. They sell it in order to cure tropical fish. As far as I know gentamycin is the best since you can pressure cook it. Here is a link that describes some other antibiotics and their effect on bacterias.


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InvisibleG a n j a
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Re: Agar recipe for cloning oysters [Re: ragadinks]
    #2174291 - 12/13/03 08:21 AM (18 years, 1 month ago)

Thank you, good link there :cool:
Now I just need to track some down, luckily theres a large farm shop with a big vet section near me, I'll try there first. Gentamycin seems too be the best alrounder.


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Offline284_27
Indocybin

Registered: 07/13/03
Posts: 193
Last seen: 6 months, 18 days
Re: Agar recipe for cloning oysters [Re: G a n j a]
    #2174353 - 12/13/03 10:09 AM (18 years, 1 month ago)

am not trying to hijack the thread , but how what ratio of gentamycin is added ?


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"I find that the harder I work, the more luck I seem to have."
- Thomas Jefferson

Fly high and I will meet you there .
- Timothy Leary


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Offlineragadinks
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Re: Agar recipe for cloning oysters [Re: 284_27]
    #2174371 - 12/13/03 10:39 AM (18 years, 1 month ago)

I think it is 100 mg per liter. Have a look at this thread.


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Offlinepluteus
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Registered: 08/12/03
Posts: 170
Loc: London area, UK
Last seen: 15 years, 2 months
Re: Agar recipe for cloning oysters [Re: ragadinks]
    #2174727 - 12/13/03 03:00 PM (18 years, 1 month ago)

I add at 100mg/litre. I have several sources for gentamycin sulphate here in the UK, but it's quite an expensive substance - around ?140 for 5 grams of 60% purity = 50 litres of antibiotic agar media at ?2.80 each

I find that it never seems to eradicate bacteria at this concentration, just - when it works - reduces them to maneagable levels (little blobs as opposed to big splodges) so that it's generally much easier to isolate clones.

I've got a friend who uses novobiocin with success, but I can't remember if it's autoclavable. It is, however, much cheaper. As regards autoclavability, I don't think antibiotic carefully added to cooling media really constitutes a serious vector of contamination anyway, definitely not in my experience.

Gentamycin works by disabling part of the cell machinery (a ribosome protein) that makes other proteins. It does this in a range of organisms, including - according to my jar of it - Gram-negative and Gram-positive bacteria, and mycoplasma, but - crucially - not basidiomycetes. Resistance is commonplace, so it is really just another tool and not a cure-all.


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InvisibleWheedhead!
Test subject #1

Registered: 11/14/02
Posts: 205
Loc: Area 51
Re: Agar recipe for cloning oysters "UPDATE GROWTH" [Re: Wheedhead!]
    #2183052 - 12/16/03 05:43 PM (18 years, 1 month ago)

Ok, so I finally moved my lazy ass to my friends house and got my camera.  I just thought I ould share some pics of the above mentioned projekt.

These Plates were all made 22-11-03. 













2 plates has some yellow on them, it looks like mycelium piss to me? Anyway ill just use pieces without it.
Tomorrow ill transfer some vedges to corn.

Enjoy the pics  :cool:


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InvisibleG a n j a
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Registered: 12/03/02
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Loc: Zone ate
Re: Agar recipe for cloning oysters "UPDATE GROWTH" [Re: Wheedhead!]
    #2183298 - 12/16/03 06:53 PM (18 years, 1 month ago)

nice pics, good job :thumbup:
That is more than likely mycelium piss as i see it
far more in oysters than cubensis.


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