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oldmanofthewoods



Registered: 08/10/10
Posts: 274
Loc: Pennsylvania
Last seen: 3 years, 9 months
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Spore print to agar- how many transfers?
#16000758 - 03/26/12 08:15 PM (11 years, 10 months ago) |
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I have a few oyster prints that I tried growing on agar. I made about 15 plates with three different varieties. All of the plates were contaminated with a blue/green mold. Disappointed, I threw most of them away. I did keep 4 of the less moldy plates with evidence of mycelium growing and transfered again. These new plates were just as moldy and some of the mold spots were outside the place I rubbed the inoculation loop.
How many times do you typically transfer from plates to get a pure culture?
Also, is the mold growing all over the surface of the agar a sign of air-contamination? I should mention I did a few clones on agar that same day and 4/5 of them are contaminate free..which makes me think it's not an air problem..
I still have some prints of varieties that I don't have a fresh specimen to clone, so I would like to know what's going on with my moldy plates. Any advise will be greatly appreciated. Thanks!
-------------------- A collection of my mushroom photos > fungi of pennsylvania A giant puffball can produce 7-9 trillion spores. If each spore produced one giant puffball they would reach to the sun and back. - David Arora, Mushrooms Demystified
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cc2
Mush

Registered: 05/15/10
Posts: 2,611
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Quote:
oldmanofthewoods said: How many times do you typically transfer from plates to get a pure culture?
as many as are needed to get a satisfactory result, nothing but mycelium growing on agar. 
Quote:
oldmanofthewoods said: Also, is the mold growing all over the surface of the agar a sign of air-contamination?
it could be, but since you transferred from moldy plates, your inoculation loop most probably carried some mold spores along with the isolated mycelium.
also working in a glove box or even better under a flow hood helps immensely.
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oldmanofthewoods



Registered: 08/10/10
Posts: 274
Loc: Pennsylvania
Last seen: 3 years, 9 months
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Re: Spore print to agar- how many transfers? [Re: cc2]
#16007119 - 03/28/12 06:42 AM (11 years, 10 months ago) |
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I'm looking for some kind of magic number. I only have 20 small half pint mason jars and I 20 plastic petri dishes. If it takes several transfers (7-10? more?) I don't think my current set up can handle that many.
I do have a glove box. I might do some reading on antibacterial stuff people add to their agar. I briefly read about that in the past, but I never had this much mold on my agar plates.
-------------------- A collection of my mushroom photos > fungi of pennsylvania A giant puffball can produce 7-9 trillion spores. If each spore produced one giant puffball they would reach to the sun and back. - David Arora, Mushrooms Demystified
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cc2
Mush

Registered: 05/15/10
Posts: 2,611
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Quote:
oldmanofthewoods said: I'm looking for some kind of magic number. I only have 20 small half pint mason jars and I 20 plastic petri dishes. If it takes several transfers (7-10? more?) I don't think my current set up can handle that many.
there are no magic numbers besides π and e, if any.
if you have fewer supplies than your projects need, simply hold a few of them off there ain't going to be any breakthrough solutions other than getting more supplies or holding off some strains.
Quote:
oldmanofthewoods said: I do have a glove box. I might do some reading on antibacterial stuff people add to their agar. I briefly read about that in the past, but I never had this much mold on my agar plates.
antibacterial compounds, as the name implies, inhibit reproduction and growth of bacteria colonies, not of mold ones, which are still fungi. the only thing you can do to keep molds out of your mycelium is to isolate healthy sectors and work clean until you have no mold on your plates.
unfortunately being molds and other fungi so closely related, a mold inhibitor or killer is going to affect your oyster mycelium as well.
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fr0st
Oddjob


Registered: 12/09/11
Posts: 99
Last seen: 8 years, 1 month
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Re: Spore print to agar- how many transfers? [Re: cc2]
#16008665 - 03/28/12 12:48 PM (11 years, 10 months ago) |
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Quote:
cc2 said: there are no magic numbers besides π and e, if any.
Don't forget about c
On a side note, sorry I don't have anything productive to add
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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,865
Last seen: 1 day, 18 hours
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Re: Spore print to agar- how many transfers? [Re: fr0st]
#16008733 - 03/28/12 12:59 PM (11 years, 10 months ago) |
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Could be as little as 1 transfer from the first dish could be more. Mold really isn't that hard to isolate from, at least not the ones I've dealt with. If it's a dirty print though there may be bacteria on there as well so look for that too. That's harder to isolate from, and the antibacterial agar may help with that if that's the case.
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