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 tomldp
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Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 19 years, 7 months
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 Save contaminated spores with agar ? 
#603375 - 04/09/02 01:58 PM (21 years, 1 month ago) |
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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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 Visit and support the Free Spore Ring Europe
Edited by tomldp (04/09/02 03:30 PM)
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vatoloco
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Registered: 01/30/02
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 Post deleted by MOE THE MAD SCIENTIST [Re: tomldp]
#603425 - 04/09/02 02:52 PM (21 years, 1 month ago) |
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ParticleMan
enthusiast
Registered: 02/05/02
Posts: 240
Last seen: 21 years, 1 month
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Re: Save a contaminated print on agar ? [Re: vatoloco]
#603433 - 04/09/02 03:01 PM (21 years, 1 month ago) |
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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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tomldp
Shroomeur
Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 19 years, 7 months
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Re: Save a contaminated print with agar ? [Re: vatoloco]
#603451 - 04/09/02 03:23 PM (21 years, 1 month ago) |
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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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Visit and support the Free Spore Ring Europe
Edited by tomldp (04/09/02 03:33 PM)
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 CLuB99
lost somewhere in time and space
 
Registered: 10/26/99
Posts: 1,316
Loc: my mind
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Re: Save a contaminated print with agar ? [Re: tomldp]
#603491 - 04/09/02 03:56 PM (21 years, 1 month ago) |
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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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DinoMyc
Ipsa scientiapotestas est
Registered: 11/13/99
Posts: 1,080
Last seen: 16 years, 11 months
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Re: Save contaminated spores with agar ? [Re: tomldp]
#603829 - 04/09/02 11:23 PM (21 years, 1 month ago) |
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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 affront, I apologize.
If I made affirmation, I apologize.
I merely came to listen, came to say.
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tomldp
Shroomeur
Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 19 years, 7 months
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Re: Save contaminated spores with agar ? [Re: DinoMyc]
#603918 - 04/10/02 01:14 AM (21 years, 1 month ago) |
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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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Visit and support the Free Spore Ring Europe
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Seuss
Error: divide byzero
Registered: 04/27/01
Posts: 23,480
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Re: Save contaminated spores with agar ? [Re: tomldp]
#604179 - 04/10/02 10:12 AM (21 years, 1 month ago) |
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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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tomldp
Shroomeur
Registered: 10/03/01
Posts: 389
Loc: Europe
Last seen: 19 years, 7 months
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Re: Save contaminated spores with agar ? [Re: Seuss]
#604344 - 04/10/02 01:25 PM (21 years, 1 month ago) |
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I'll have a lot of job, but I should learn a lot from it. Thanx a lot guys !
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Visit and support the Free Spore Ring Europe
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ParticleMan
enthusiast
Registered: 02/05/02
Posts: 240
Last seen: 21 years, 1 month
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Re: Save contaminated spores with agar ? [Re: Seuss]
#604591 - 04/10/02 05:34 PM (21 years, 1 month ago) |
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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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