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Invisibletryptonite
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PDA help
    #2573224 - 04/18/04 07:12 AM (12 years, 7 months ago)

I started 10 cultures on PDA plates by spore prints about 5 days ago. This plate is the best one I think (has the whitest mycelium). Is this sufficient to put on my substrate yet? Why doesnt it look all spiraly like other mycelium? What should i do next?



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Offlineragadinks
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Re: PDA help [Re: tryptonite]
    #2573239 - 04/18/04 07:38 AM (12 years, 7 months ago)

I would isolate the best looking bit of mycelium to another plate.
It's hard to tell from the picture, but it seems to me that you have some contamination on the plate.
If that is true you would have to isolate the best looking mycelium away from the contamination.


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Invisibletryptonite
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Re: PDA help [Re: ragadinks]
    #2575948 - 04/19/04 12:46 AM (12 years, 7 months ago)

so the thick, bright white mycelium is the one I have to isolate?


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OfflinePsilygirl
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Re: PDA help [Re: tryptonite]
    #2576140 - 04/19/04 01:34 AM (12 years, 7 months ago)

it kinda looks like you have some contam there... some bacteria most likely.

did you put the petri in the bottom of your incubator? try propping it up on something like styrafoam half an inch or so, this got rid of all my petri bacterial contams... seems it laying directly on the incubator floor was too warm.

try this on your next petris and see if it helps.


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Invisibletryptonite
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Re: PDA help [Re: Psilygirl]
    #2576739 - 04/19/04 06:19 AM (12 years, 7 months ago)

Quote:

Psilygirl said:
did you put the petri in the bottom of your incubator?




> dont have an incubator.

Would it be ok to isolate the good mycelium onto new plates or will it carry the infection?


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InvisibleLoki
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Re: PDA help [Re: tryptonite]
    #2576802 - 04/19/04 07:31 AM (12 years, 7 months ago)

You should be able to transplant a good section, place it upside down on the fresh plate, then transphere again and new growth you get, nice and early, so as to hopefully leave the contam behind.

One reason to flip it is, if you are transplanting to pda + peroxide, the
contam spores will be ontop of the mycelium, so if you transplant the
mycelium face down, onto the pda + p you will have a better chance of leaving behind the contam.


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Invisibletryptonite
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Re: PDA help [Re: Loki]
    #2576968 - 04/19/04 09:12 AM (12 years, 7 months ago)

Quote:

Loki said:
You should be able to transplant a good section, place it upside down on the fresh plate, then transphere again and new growth you get, nice and early, so as to hopefully leave the contam behind.




>What do you mean? Are you talking abouting cutting a chunk off and transferring to another dish? I just re-innoculated the white mycelium using a loop.

Quote:

Loki said:
One reason to flip it is, if you are transplanting to pda + peroxide, the contam spores will be ontop of the mycelium, so if you transplant the mycelium face down, onto the pda + p you will have a better chance of leaving behind the contam.




> My dishes are always upside down so maybe the contam spores are on the lid? Also how do I use peroxide on the plates? :wexican:


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Invisiblesakura
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Re: PDA help [Re: tryptonite]
    #2576991 - 04/19/04 09:33 AM (12 years, 7 months ago)

In these two volumes is all you need to know about working with h2o2 (from the man who pioneered the technique :wink: )

http://www.nansnook.com/~archives/tek/Volume1.html


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InvisibleLoki
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Re: PDA help [Re: tryptonite]
    #2577248 - 04/19/04 11:33 AM (12 years, 7 months ago)

Yes, I mean to cut out a good peice of mycelium and agar, and transphere it to a new petri dish of pda+h2o2

Sakuras link is good here is some more
reading to do on agar + peroxide

http://www.shroomery.org/index/par/23462

I dont transphere with a loop, I only use a loop to
start spores, every other transpher i make i make by
cuting a chunk of mycelium and agar from the mother plate
and move it onto its own fresh plate containing peroxide.

You cannot start spores on peroxidated agar, only transphere
existing mycelium.


As to the lids and storing your plates upside down, you
miss my point i think.

My point is that if there is a contam in the plate, the whole surface of the
agar could harbour spores, or endo spores from the contam, so
If when you transphere a chunk, you place it mycelium side down, and
spores or endo spores will not germinate, as they are in direct
contact with the peroxidated agar, and only the existing mycelium will
be able to spread.


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Invisibletryptonite
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Re: PDA help [Re: Loki]
    #2581330 - 04/20/04 04:02 AM (12 years, 7 months ago)

ah I understand now. Thanks for the advice!


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