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Neowynd8

Registered: 06/12/16
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Lc test time frame
#23626899 - 09/08/16 07:12 PM (7 years, 4 months ago) |
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I prepared a honey lc solution and inoculated with Ms syringe. It has been colonizing for approx. 3 weeks. I inoculated 3 standard pk tek brf jars in order to discern if the lc is contaminant-free or not. The test jars displayed growth in under 24 hours. From my understanding, extremely fast growth can be indicative of contamination, but I'm confident that only healthy mycelium is present as of now. I intend on inoculating more lc with this one in order to expand my stock. How long/much should I allow the test jars to colonize before confirming a contaminant-free lc?



Edited by Neowynd8 (09/08/16 07:21 PM)
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bodhisatta 
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Registered: 04/30/13
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Re: Lc test time frame [Re: Neowynd8]
#23626975 - 09/08/16 07:30 PM (7 years, 4 months ago) |
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you should get fast growth from LC since it's active mycelium. ideally you use it at the peak of growth phase because after it eats all the nutrients from the LC it will start to deplete it's energy reserve(mostly glycogen).
testing it on a cake is fine. you just need an eye for cubensis mycelium. otherwise you can wait and fruit it and see that way.
ideally you just use the spores right in a PF cake. since it's faster and less steps(which means one less chance for contamination) and less work to just use spores right to a grow.
you have to wait for the LC to colonize and then wait to test it you would have been quite further ahead had you just used the spores. and since mushrooms make more spores you don't have to worry about using your whole syringe up. so it's not like you're saving spores by turning them into LC.
https://www.shroomery.org/forums/showflat.php/Number/21365919#21365919
there's too many spores in a syringe as the above link shows. as you put spores in a LC even if there's 1 drop. you put in thousands of spores. say there's 1 contaminant(bacteria or spore) per 10,000 spores. you get even 1 contaminant in a LC and it's garbage. if you squirt spores in a PF cake the contaminants are slightly mitigated because of the way PF cake mix works. sometimes a grow gets fucked but largely the inherent contaminants in a spore syringe are not a huge deal on cakes.
ideally you make a LC with a clean culture on agar. you start the spores on agar then make the LC.
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Neowynd8

Registered: 06/12/16
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Surely it isn't necessary to proceed entirely through the fruiting stage to confirm a clean lc....
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bodhisatta 
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Re: Lc test time frame [Re: Neowynd8]
#23627308 - 09/08/16 09:34 PM (7 years, 4 months ago) |
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Unless you can't tell clean mycelium on agar or a cake
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PsilocyBen17
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Re: Lc test time frame [Re: bodhisatta] 1
#23627480 - 09/08/16 10:30 PM (7 years, 4 months ago) |
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Spores to LC is one of the worst ideas that gets brought up on here...
Spores are almost always full of contams since they are never harvested under sterile conditions...if you need to make LCs its a much better idea to go spores>agar>LC to ensure cleanliness...
I'd be weary of growth after 24 hours too. Could be trich or anything really. Do your jars have a dry verm layer? they look pretty full to me.
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Neowynd8

Registered: 06/12/16
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Yes, they have a dry verm layer. I realize ms to lc isn't ideal. I'm actually attempting various methods of expansion...ms to agar made from pre mixed powder I ordered online, which is an ongoing disaster for various reasons and grain lc. I suppose it would be best to inoculate these larger quart jar lcs with the grain lc from this corn huh?

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PsilocyBen17
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Re: Lc test time frame [Re: Neowynd8]
#23627633 - 09/08/16 11:52 PM (7 years, 4 months ago) |
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Why not just g2g if you are looking at expanding your spawn? Or make a liquid inoculate with the colonized jar? alot more vectors doing it any other way...
Can I ask why your agar is a disaster?
Edited by PsilocyBen17 (09/09/16 12:04 AM)
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Neowynd8

Registered: 06/12/16
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Last seen: 7 months, 2 days
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G2G seemed risky. The plan has always been to make liquid inoculant. I just pced a pint jar of water for it. The agar wouldnt dissolve despite vigorous boiling and gradual addition to water. Alot of it would but not all. So I had concerns about it not being sufficiently concentrated. I ended up pouring it into half pint jars then pcing...I realized afterward that with using half pint jars with no coloring agent,as opposed to petri dishes, would make it incredibly difficult to visually inspect the agar. Perhaps that's only because there isn't any perceptible mycelium yet lol...I inoculated them with ms.
Edited by Neowynd8 (09/09/16 12:08 AM)
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bodhisatta 
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Re: Lc test time frame [Re: Neowynd8]
#23628281 - 09/09/16 07:47 AM (7 years, 4 months ago) |
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G2G is far less risk than LC if you build a still air box
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Neowynd8

Registered: 06/12/16
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All my endeavors are conducted in a SAB. G2G seems riskier than making a liquid inoculant with sterilized water.
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PsilocyBen17
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Re: Lc test time frame [Re: Neowynd8]
#23628798 - 09/09/16 11:46 AM (7 years, 4 months ago) |
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Can you explain how?
A LI involves an extra vector of water...
More vectors=riskier procedures.
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bodhisatta 
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And needles are horribly nooby technique. Using injection ports too.
I still use both. But realise they're not great
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Mushierage
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Registered: 06/29/16
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Quote:
Trusted Cultivator said:

And needles are horribly nooby technique. Using injection ports too.
I still use both. But realise they're not great
You mean for drawing up LC?
I find that putting a ship in a pasty plate works really well for drawing up LI. Pour methods may be better due to less contam vectors though.
-------------------- Don't like researching posts? Read this! . Also, if you're new and your posts contain the words: Humidifer, incubator, air-stone, or heater, then you need to read and UTFSE before asking people to review your setup. OR... You should be cultivating reptiles and fish, not mushrooms.
  
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Neowynd8

Registered: 06/12/16
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Last seen: 7 months, 2 days
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Quote:
PsilocyBen17 said: Can you explain how?
A LI involves an extra vector of water...
More vectors=riskier procedures.
Exposure to open air? Despite being in SAB.
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