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Invisibletahoe
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Transferring contams away from the mycelium
    #4100718 - 04/26/05 11:35 PM (11 years, 7 months ago)

i had a much needed culture that contaminated. This Culture was isolated once from spores and i didn't want to go any further then that so i could keep the diversity very high. Plus i was able to get good rhizos from this strain that usually doesn't go rhizomorphic on agar. So i cut the contams out, these were contams due to my shitty glove box and were not brought in my the mushrooms spores. There were 2 spots, they were not allowed to get bigger then a small pea. So do you guys think this culture is good enough for lc? Anyone ever transfer this way with luck? Here it is, one contam was found early and is that small u shaped area right about the word isolate. The second contam was at the edge so i just cut out the remaining uncolonized agar so that the mycelia would not have any more contams


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Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


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InvisibleOlgualion
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4100840 - 04/27/05 12:25 AM (11 years, 7 months ago)

I'd recommend transferring to another plate first to be sure the mycelium is not harboring mold spores. If using H2O2, be sure that the final piece used for inoculating your malt water is not touching the inital transfer. You may also get away with either dipping a wedge (directly from that plate) in H2O2, or by adding peroxide to your liquid malt solution, then transferring from that plate. IMO the first is the "better practice".

But I am no mycologist, and others' opinions may vary...


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Invisibletahoe
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Re: Transferring contams away from the mycelium [Re: Olgualion]
    #4100846 - 04/27/05 12:27 AM (11 years, 7 months ago)

i need the genetic diversity of this petri. I dont really want to go to another dish


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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Offlinestarspawn2
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4101367 - 04/27/05 03:02 AM (11 years, 7 months ago)

Quote:

i need the genetic diversity of this petri


In the picture it looks like a piece of a mushroom is in the middle of the petri dish,is that an isolate?If so what "genetic diversity" do you mean? About trying to clean your culture,always transfer the mycelium away from the contam into a new dish,not the other way around.Trying to cut the bad part out is not the best way to save it.


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InvisibleWaylitJim
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Re: Transferring contams away from the mycelium [Re: starspawn2]
    #4101924 - 04/27/05 09:34 AM (11 years, 7 months ago)

You could try the hot agar tek....
Just pour a fresh layer of agar over that dish.
Any contams will die but the mycelium will not.
This works try it...the myc will grow thru the new layer
free from contamination.


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Offlinedebianlinux
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Re: Transferring contams away from the mycelium [Re: starspawn2]
    #4102167 - 04/27/05 11:39 AM (11 years, 7 months ago)

Quote:

starspawn2 said:
In the picture it looks like a piece of a mushroom is in the middle of the petri dish,is that an isolate?If so what "genetic diversity" do you mean? About trying to clean your culture,always transfer the mycelium away from the contam into a new dish,not the other way around.Trying to cut the bad part out is not the best way to save it.



yes yes yes
stay away from contams on plates. the more you rattle them around the more of a mess (microscopically speaking) you are making.

i'm missing the whole point about "genetic diversity" as well; assuming that is an isolate. based on what little information was provided, I'd say he started from spores and is in the process of isolating (that being a wedge then). The pictured plate looks divergent to me. i take it he is trying to save the more rhizo growth that has it's ass covered in contams.

as a general rule of competence you should never try to mass propagate an even remotley suspect culture. make damn sure (via multiple transfers) that the culture is clean before trying to generate bulk amounts of spawn from it!


Edited by debianlinux (04/27/05 03:39 PM)


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Invisibletahoe
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Re: Transferring contams away from the mycelium [Re: debianlinux]
    #4102804 - 04/27/05 03:17 PM (11 years, 7 months ago)

this was isolated from a dish that had the spores on it. I believe that the more you isolate= less genetic diversity. If you isolate too many times then you run the risk of isolating a fruitless strain. This culture is for an outdoor bed and is not native to this area. This means that my climate is not perfect for this species so i dont want to isolate it down to something that wont fruit here. I know its a long shot to get it to this point but the more diverse this strain is the better chance i have of it fruiting


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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Offlinedebianlinux
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4102864 - 04/27/05 03:37 PM (11 years, 7 months ago)

Quote:

tahoe said:
I believe that the more you isolate= less genetic diversity. If you isolate too many times then you run the risk of isolating a fruitless strain.




an isolate is just that, an isolated "strain".

genetic diversity refers to what you get from a multi-spore print (the exact opposite of an isolate) You could be referring to the ability of isolated mycelium to utilise a variety of substrates. This definition actually has a proper term that escapes me atm. I know the end result or symptom of this phenomena is senescence.

You can bet that if your mycelia is still diverging on the plate then you do not, in fact, have an isolate.

You always run the risk of isolating a non-fruiting strain as this is one of the possible disadvantages of genetic diversity. Of the thousands of pairs there are, statistically speaking, weak and non-fruiting pairs. In the wild, these pairs get obsoleted in the great substrate race. On agar you are selectively weeding out pairs until you get to 1 strain. Therefore you could, theoretically, weed out all good "strains" and be left with a crappy one. You can definitely tilt these odds very far in your favor by following the basic, and obvious, selection criteria (rhizo, not tomentose, etc) laid out in many books and guides.

All this tells me that it is probably best to start multispore in a liquid culture and as the mycelium consumes inoculated substrate, well, may the best pair win. Once you have achieved fruitings from a variety of multispore grows you can get an idea of which "strain" worked out the best in the conditions you are providing. Then clone a fruit from this batch onto agar. By cloning the flesh you are starting with an isolate of a "strain" known to perform well in the conditions you are providing.

Hope this all makes sense.


Edited by debianlinux (04/27/05 03:44 PM)


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Invisibletahoe
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Re: Transferring contams away from the mycelium [Re: debianlinux]
    #4102913 - 04/27/05 03:55 PM (11 years, 7 months ago)

yes it all makes sense and i already knew it. The point was cutting away the contams but we got ot his point.
Okay which one would you rathing use for an outdoor bed?


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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Offlinedebianlinux
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4102923 - 04/27/05 03:58 PM (11 years, 7 months ago)

erm, why not both? clone from what does best.


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Offlinedebianlinux
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4102935 - 04/27/05 04:01 PM (11 years, 7 months ago)

Quote:

tahoe said:
yes it all makes sense and i already knew it.




after re reading the thread i can better understand your original query. it was the misuse of the term isolate that was throwing me.


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OfflineMushroomFriend
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Re: Transferring contams away from the mycelium [Re: debianlinux]
    #4103092 - 04/27/05 04:44 PM (11 years, 7 months ago)

Tahoe maybe you mean that transfer after transfer degenerates the mycelium?
Like when you do to many g2g one can get mutants and such?

I always learned that its better to transfer healthy, non contammed, mycelium to another plate....

MF


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Invisibletahoe
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Re: Transferring contams away from the mycelium [Re: debianlinux]
    #4103345 - 04/27/05 05:33 PM (11 years, 7 months ago)

what does one call it when you cut an area from one petri and put it to another


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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Invisibletahoe
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4103360 - 04/27/05 05:36 PM (11 years, 7 months ago)

how does one isolate then?


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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Offlinescatmanrav
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4103394 - 04/27/05 05:42 PM (11 years, 7 months ago)

To be an isolate, youd need to transfer it until there was only one mycelium. I dont know what the steps in between are called...I understood what you meant.

IMO, from grain jars with little tiny pin sized green spots at the bottom, take a few peices of grain from the top and put it to a petri, and it contams...spores throughout the whole thing from that. I'd transfer to a new plate rather then use the plate that had green on it. Who knows though. I say use both.


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"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

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Invisibletahoe
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Re: Transferring contams away from the mycelium [Re: scatmanrav]
    #4103418 - 04/27/05 05:45 PM (11 years, 7 months ago)

heres what i think. Every time you transfer you are isolating, you will be isolating untill you are down to 1 mycelium and then you would be cloning


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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Offlinedebianlinux
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Re: Transferring contams away from the mycelium [Re: tahoe]
    #4103466 - 04/27/05 05:57 PM (11 years, 7 months ago)

Quote:

tahoe said:
heres what i think. Every time you transfer you are isolating, you will be isolating untill you are down to 1 mycelium and then you would be cloning



very good point.


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Re: Transferring contams away from the mycelium [Re: debianlinux]
    #4106161 - 04/28/05 10:29 AM (11 years, 7 months ago)

agreeing with earlier posters, dont try to cut out contam. however you can try to contain it with a strong h2o2agar-sandwich just on the contam.

or, not touching the contam. cut out a separation between the two, and fill the separation with strong h2o2agar.

you could even combine the two methods.


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Re: Transferring contams away from the mycelium [Re: Mindzpore]
    #4106304 - 04/28/05 11:24 AM (11 years, 7 months ago)

Oh I forgot about the hot agar thing...I agree with that idea. Some warm agar poured over the top...according to RR, the myc comes up through, but contams stay behind. I'm not sure if thats only true of Cubie myc though...


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Re: Transferring contams away from the mycelium [Re: Mindzpore]
    #4106318 - 04/28/05 11:29 AM (11 years, 7 months ago)

Never disturb a contaminant in a petri dish. To do so simply dumps millions of contam spores all over your good mycelium. The proper procedure is to cut a very small piece of good mycelium and transfer it to a new petri dish. Do this in a glove box and not in front of a flow hood. A flow hood will blow the contaminant spores everywhere. It takes many transfers away from the original spores to get an isolated strain. You have nothing to fear from transferring away from contamination. By the time a petri dish is grown out like that one, clamp connections have formed everywhere and the genetic information has already been exchanged throughout the entire dish.
RR


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