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LC contam detection?


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#1 vitae1234

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Posted 13 November 2007 - 06:06 PM

I'm wondering if it's possible to detect contams in a LC, or if I should just step up to agar?

Reason being, I have some vendor syringes that are 2 for 2 growing trich. The first one was lucky... it also grew robust myc that obliterated the trich and ended up with successful cakes, but the second syringe was a total failure.

With 3 syringes left, I don't want to keep knocking up good substrate only to be disappointed again.

#2 prism

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Posted 13 November 2007 - 06:12 PM

LC is the way to go, Go to the archive and do some research and you can't miss.

Good luck!:rasta:

#3 Dr_T

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Posted 13 November 2007 - 06:16 PM

LC is great for growing mycelium- including mold mycelium.

If you have a syringe that is suspected of contamination, an agar plate might help you isolate a clean culture. But it's a matter of your technique, then you won't see much difference.

I'd suggest trying to take a successful jar, and start an LC from there. At least then, you have some idea what you are working with. Once you have a clean culture, LC is a powerful tool- it's worth the time it takes to do it right.

#4 prism

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Posted 13 November 2007 - 06:17 PM

Agreed!

#5 vitae1234

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Posted 14 November 2007 - 12:55 PM

Did some digging, found a thread that answers my question.

Mycotopia Web Archive: In Liquid Culture

#6 vitae1234

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Posted 06 December 2007 - 09:03 PM

FOAF ended up with 100% colonization....

... of what appears to be cobweb.

First pic: nocced with LC grown out from vendor syringe, it *looked* like myc in the LC, anyway.

Second pic (for comparison) is WBS prepped exactly the same way but nocced with a standard syringe of spores from a print FOAF took herself.

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#7 Birdman

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Posted 06 December 2007 - 10:39 PM

I learned the hard way and had to clean up quite
a mess.Now I take a pint or half pint jar of cheep millet that
was PC'd and left to sit for 7 to 10 days to be sure it's
clean.Then hit it with 1 cc and let it go to see what happends.
If its trec it will go off and with in three days it will be green.
If after a week it's still clean I use a couple cc's to make LC
and knock up the rest of the jars.
This is what bad LC on millet looks like and after only three days.
The man stood good for it though but set me back a week or so.
Will see how the next jar goes.

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#8 MePlusOne

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Posted 07 December 2007 - 02:58 AM

I don't know if anyone else has ever done this, but if I think an LC is in question, then when I draw it out of the LC with a clean syringe, I inject that into a small shot glass of hydrogen peroxide and water and let it sit for a minute and draw it back into another clean syringe (all while in a sterile environment of course)

Eventualy the peroxide breaks down to water anyway and I have had great results with this way of doing things. that's just my experience though. I will admit that not every LC I did this with always kept it from contaming, but more survived than were lost. It works in my opinion.
But the mycelia does suffer somewhat of a "burn" from the rinse. I've noticed some LCs losing their vigor more quickly than others but there could be other variables at play too or in combination with this.

Also, I believe that too much peroxide and you will suffer pink/red mold, but that is just my theory through from my experience...

#9 Hippie3

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Posted 07 December 2007 - 12:24 PM

interesting if a bit burdensome,
lc is so easy to get a large amount
might not be worth that 'heroic' measure .
i suspect mixing with a 200:1 water:bleach solution would also help in cleaning up mildly contaminated lc solution before use.
some good contam pix & info herein

#10 MePlusOne

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Posted 07 December 2007 - 03:41 PM

I know it may be a little over-the-top, but I just like to be somewhat assured before I use my LCs cause I hate to use one that I have been waiting on developing and then finding out it is contamed =(

This is just something that seems to work for me, but as you said, bleach may be more efficient for this.

#11 Hippie3

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Posted 07 December 2007 - 04:09 PM

the best approach, imo , is to test-fire any lc on one test jar before using it inoculate dozens.

#12 MePlusOne

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Posted 07 December 2007 - 04:17 PM

very rarely, but occasionally I will just put like an inch of brf/verm in a half-pint with a silicone injector site and just use a small amount of LC and see if it will colonize that without contaming, but I do it only in the most questionable instances.

Hippie is right, a test jar is the best way overall, but there are other methods that work without using this one imo.

Peace.




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