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pluteus
level-9 deviant
Registered: 08/12/03
Posts: 170
Loc: London area, UK
Last seen: 17 years, 4 months
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Re: Agar recipe for cloning oysters [Re: G a n j a]
#2172061 - 12/12/03 01:38 AM (20 years, 3 months ago) |
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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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G a n j a
Pictish and proud
Registered: 12/03/02
Posts: 7,860
Loc: Zone ate
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Re: Agar recipe for cloning oysters [Re: pluteus]
#2172107 - 12/12/03 02:05 AM (20 years, 3 months ago) |
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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 Do antibiotic's stop bacteria ?
-------------------- er
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ragadinks
MrBeatle
Registered: 10/20/03
Posts: 1,298
Last seen: 5 months, 18 days
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Re: Agar recipe for cloning oysters [Re: G a n j a]
#2173352 - 12/12/03 04:45 PM (20 years, 3 months ago) |
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I do not know what the situation is like in the UK. I got gentamycin from my local doc ( but he's a relative ... ). 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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G a n j a
Pictish and proud
Registered: 12/03/02
Posts: 7,860
Loc: Zone ate
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Re: Agar recipe for cloning oysters [Re: ragadinks]
#2174291 - 12/13/03 06:21 AM (20 years, 3 months ago) |
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Thank you, good link there 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.
-------------------- er
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284_27
Indocybin
Registered: 07/13/03
Posts: 193
Last seen: 2 years, 8 months
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Re: Agar recipe for cloning oysters [Re: G a n j a]
#2174353 - 12/13/03 08:09 AM (20 years, 3 months ago) |
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am not trying to hijack the thread , but how what ratio of gentamycin is added ?
-------------------- "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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ragadinks
MrBeatle
Registered: 10/20/03
Posts: 1,298
Last seen: 5 months, 18 days
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Re: Agar recipe for cloning oysters [Re: 284_27]
#2174371 - 12/13/03 08:39 AM (20 years, 3 months ago) |
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I think it is 100 mg per liter. Have a look at this thread.
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pluteus
level-9 deviant
Registered: 08/12/03
Posts: 170
Loc: London area, UK
Last seen: 17 years, 4 months
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Re: Agar recipe for cloning oysters [Re: ragadinks]
#2174727 - 12/13/03 01:00 PM (20 years, 3 months ago) |
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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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Wheedhead!
Test subject #1
Registered: 11/14/02
Posts: 205
Loc: Area 51
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Re: Agar recipe for cloning oysters "UPDATE GROWTH" [Re: Wheedhead!]
#2183052 - 12/16/03 03:43 PM (20 years, 3 months ago) |
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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
-------------------- I just The Shroomery Monkeys might fly out of my butt Dope will get you through times of no money better than money will get you through times of no dope. " Fabulous Furry Freak Brothers
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G a n j a
Pictish and proud
Registered: 12/03/02
Posts: 7,860
Loc: Zone ate
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Re: Agar recipe for cloning oysters "UPDATE GROWTH" [Re: Wheedhead!]
#2183298 - 12/16/03 04:53 PM (20 years, 3 months ago) |
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nice pics, good job That is more than likely mycelium piss as i see it far more in oysters than cubensis.
-------------------- er
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