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Offlinetomldp
Shroomeur

Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 20 years, 7 months
Save contaminated spores with agar ?
    #603375 - 04/09/02 01:58 PM (22 years, 2 months ago)

I received a heavily contaminated and already used print of mexicub (I made a syringue and inoculated two jars with these spores). I want to isolate an uncontaminated piece of mycelium so that I can grow this strain.
Do I have a chance ? There is at least 3 different contaminations growing along with the mycelium.
I thought that by selecting each time a "clean" piece of mycelium, I should manage to isolate good mycelium in order to grow this strain.
What should I try if I want to get a clean piece of mycelium ? Should I start my work by injecting spores on agar (= starting from the very beginning), or cut a piece of white mycelium from a contaminated jar and put it on agar (= using what I already have) ?


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Edited by tomldp (04/09/02 03:30 PM)

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Offlinevatoloco
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Registered: 01/30/02
Posts: 7,653
Last seen: 19 years, 8 months
Post deleted by MOE THE MAD SCIENTIST [Re: tomldp]
    #603425 - 04/09/02 02:52 PM (22 years, 2 months ago)


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OfflineParticleMan
enthusiast
Registered: 02/05/02
Posts: 240
Last seen: 22 years, 1 month
Re: Save a contaminated print on agar ? [Re: vatoloco]
    #603433 - 04/09/02 03:01 PM (22 years, 2 months ago)

what i have found so far with agar is that it is much easier to clone tissue than to do a multi-spore, especially if contams are involved. if you are going to try agar i would suggest cloning a good part of the myc.


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

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Offlinetomldp
Shroomeur

Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 20 years, 7 months
Re: Save a contaminated print with agar ? [Re: vatoloco]
    #603451 - 04/09/02 03:23 PM (22 years, 2 months ago)

I should have been more precise (changed my initial post).

I think I'll try to cut a piece of mycelium then "purify" it. If I'm courageous enough, I'll post pics of that work.

Thanx for your advices !


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:laugh: Visit and support the Free Spore Ring Europe :laugh:

Edited by tomldp (04/09/02 03:33 PM)

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InvisibleCLuB99
lost somewhere in time and space
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Registered: 10/26/99
Posts: 1,316
Loc: my mind
Re: Save a contaminated print with agar ? [Re: tomldp]
    #603491 - 04/09/02 03:56 PM (22 years, 2 months ago)

clone the mycelium you already have on a peroxi agar plate, then try to clean it sectoring again until pure culture is obtained

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OfflineDinoMyc
Ipsa scientiapotestas est
Registered: 11/13/99
Posts: 1,080
Last seen: 18 years, 5 days
Re: Save contaminated spores with agar ? [Re: tomldp]
    #603829 - 04/09/02 11:23 PM (22 years, 2 months ago)

what 3 contams?
it is strongly recommended to be VERY careful with some contams.. and with 3? it sounds like you have alot of problems somewhere along the line..
you can use agar for the spores.. try it..
i would leave the contam mycelium in the trash, complete with jar.. that or sterialize the jar and toss its contents post steambath...


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If I made affirmation, I apologize.
I merely came to listen, came to say.

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Offlinetomldp
Shroomeur

Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 20 years, 7 months
Re: Save contaminated spores with agar ? [Re: DinoMyc]
    #603918 - 04/10/02 01:14 AM (22 years, 2 months ago)

I'll post a pic in a few days. I never had contams before I receive this print. The print was already used and I could hardly make one syringue from it. And I got 3 contams !!! I would call that a bad trade...


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:laugh: Visit and support the Free Spore Ring Europe :laugh:

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OfflineSeussA
Error: divide byzero

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Registered: 04/27/01
Posts: 23,480
Loc: Caribbean
Last seen: 4 months, 3 days
Re: Save contaminated spores with agar ? [Re: tomldp]
    #604179 - 04/10/02 10:12 AM (22 years, 2 months ago)

A couple of things to remember. If your contamination is spore forming, you have to be very careful to transfer the agar wedge before the spores form. If you already have a lot of spores, be sure you turn off your flow hood first for a few minutes or the turbulance will distribute the spores (assuming you have a postive pressure hood). Place the wedge mycelium down on the new plate, rather than up... again to help prevent spores from becoming airborn. Use an antibiotical agar to get rid of bacterial contaminates. Don't be afraid to play with the temperature, humditiy, and CO2 ratios. If you can identify your contaminations you maybe able to alter the environment enough to kill the contaminate but not the mycelium. Do lots of transfers sooner rather than a few transfers later. Once you think you have a clean plate, be sure to continue and grow out a few more plates just to make sure. Remember, you will be isolating a single dikaryotic growth which may not be fruit bearing. Don't throw away all your starting material... you may need it again. Once you get a fruitbody, make a clone and you are home free.

When you transfer the agar wedge, place the mycelium face down instead of up.


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Just another spore in the wind.

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Offlinetomldp
Shroomeur

Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 20 years, 7 months
Re: Save contaminated spores with agar ? [Re: Seuss]
    #604344 - 04/10/02 01:25 PM (22 years, 2 months ago)

I'll have a lot of job, but I should learn a lot from it. Thanx a lot guys !


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:laugh: Visit and support the Free Spore Ring Europe :laugh:

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OfflineParticleMan
enthusiast
Registered: 02/05/02
Posts: 240
Last seen: 22 years, 1 month
Re: Save contaminated spores with agar ? [Re: Seuss]
    #604591 - 04/10/02 05:34 PM (22 years, 2 months ago)

Does anyone have a picture of a multi-spore injection on an agar plate? My tissue clones look great (thanks to all of you helpful people) but I just had to try a multi-spore for the fun of it. When the spores germinate do they look like small white dots at first? Or is this contamination?


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

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