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invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 20 days
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LME/DEX lc experiment, how am I doing?
#19321786 - 12/24/13 03:28 PM (10 years, 1 month ago) |
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Hi,
In the past I've done honey water lcs with a fair bit of success, and since I heard LME/DEX was better I thought I should get some experience. I don't really know exactly what to expect so if you have some experience, please take a look at these pics and tell me if you think I am on the right track.
LC jar #1, injected with 10ccs of 4 month old honey water lc that was still bright white.

LC jar #2, ms inoculation from syringe that was about 6-7 months old.

LC jar #3, injected from 10ccs of 4 month old lc, different variety than jar #1.

After swirling a jar:

Another jar just after swirl:

After swirling the LC injected lcs, I can see small balls of white myc, but they generally don't get any bigger than a black pepper corn in size. I've seen LME/DEX lcs where the myc is super thick and is all one blob.
My question is, is this LC finished? Or does it always end up as a super thick blob if done right?
The MS lcs don't seem to be doing anything. The syringes were stored at ambient temps, during the summer we had a month where it was about 85 for most of the month, I wonder if those syringes are dead or what. I knocked up about 16 1/2 pint jars with 1cc of ms from more than one syringe and I haven't seen any signs of growth.
Misc info: How I prepared the lc: boiled water, added 2% powdered DEX derived from corn. Added 2% LME which was like a solid rock because I didn't know to keep it in a sealed container. I stirred until it all dissolved. I didn't filter the fluid before pcing. PC was 15 psi for 20 minutes. inoculated about 12+ hours after it was pced (let cool in the pc overnight). Lid has sfd with self healing injection port. the hole for the sfd is 3/4 inch, based on TL's grain jar tek.
Jars were inoculated on 12/21. One ms spore jar was inoculated on or around 12/17 and it looks no different from the newer ms spore jars except for one almost perfect sphere of nebulous gunk about the size of a marble. I swirl them about twice a day or whenever I think about it. I left the ms jars alone for the first day without swirling to give them a chance to grow. Temps have been 60-72 day and night.
K a big thanks to the people who contribute on this forum, you know who you are.
Edited by invitro (12/24/13 03:48 PM)
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
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Re: LME/DEX lc experiment, how am I doing? [Re: invitro]
#19321880 - 12/24/13 03:48 PM (10 years, 1 month ago) |
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i don't exactly know wut your questions are but there not always like thick n boogery i found that to happen when i didn't stir enough or used 5% and not 4 or 3
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 20 days
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Re: LME/DEX lc experiment, how am I doing? [Re: cronicr]
#19321897 - 12/24/13 03:53 PM (10 years, 1 month ago) |
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I am eager to re-inoculate the 16 1/2 pints that are not doing anything, and want to try to use one of the 'lc' lcs to accomplish this. But as I don't really know well enough what to look for I thought I'd run it by the pros in case the lc doesn't look right, or to immature or ? Plus I wanted to give a general overview of my technique because I was wondering if rock-hard LME was bad, and if maybe the 3/4 inch sfd hole was too much fae, and might somehow stunt the growth of the lc.
Edited by invitro (12/24/13 03:55 PM)
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
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Re: LME/DEX lc experiment, how am I doing? [Re: invitro]
#19321915 - 12/24/13 03:57 PM (10 years, 1 month ago) |
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y so many lc's? anyway with a good sterile technique it's not really ever too early. when i'm in doubt about the growth i stop swirling for a couple days, if it's cube myc it usually floats t the top for air where as alot of contams don't need to
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 20 days
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Re: LME/DEX lc experiment, how am I doing? [Re: cronicr]
#19321952 - 12/24/13 04:03 PM (10 years, 1 month ago) |
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I did multiple lcs this time because I only did one last time and it turned out to be 'cobweb' mold. It was really vigorously growing in the jar, like a huge blob. So I figured if I make a bunch of lcs, even if only one turns out then I'm good.
"it's not ever too early" Does this mean even if I suck up fluid from the top half of the jar, which is relatively clear, that it will be effective as an innoculant (because it has small amount of mycelium in it)? I just want to make sure the lc is going to be potent so that I'm not waiting forever on weakly inoculated grain jars when I should have let the lc mycelium grow more to begin with.
Are you saying that while the myc is growing it will stay on the bottom, but when it stops growing it floats? Or does it always float? I don't anything floating to the top yet.
Edited by invitro (12/24/13 04:05 PM)
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
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Re: LME/DEX lc experiment, how am I doing? [Re: invitro]
#19321972 - 12/24/13 04:09 PM (10 years, 1 month ago) |
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it should float if you stop swirling it, it needs air to exchange gases so it will make it's way to the surface, if it doesn't it's probly a contam
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 20 days
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Re: LME/DEX lc experiment, how am I doing? [Re: cronicr]
#19321980 - 12/24/13 04:12 PM (10 years, 1 month ago) |
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That's odd. I have done a couple dozen 1/2 pint honey water lcs to date, many of which turned out great and none of them ever floated.
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
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Re: LME/DEX lc experiment, how am I doing? [Re: invitro]
#19322061 - 12/24/13 04:16 PM (10 years, 1 month ago) |
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then it's getting enough air, this may not happen over a day or 2 but the only way to know if it's clean is to test it, thats it theres absolutely no way to tell otherwise. spores to lc is sketchy to beging with, not to say it won't or doesn't work just not the best approach. let one of your lc's colonize till your satisfied and test it. if it's clean knock more up
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 1 month, 20 days
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Re: LME/DEX lc experiment, how am I doing? [Re: cronicr]
#19322074 - 12/24/13 04:19 PM (10 years, 1 month ago) |
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Ok, getting ready to test it! Thanks again.
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newera
Stranger


Registered: 03/05/13
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Re: LME/DEX lc experiment, how am I doing? [Re: invitro]
#19322195 - 12/24/13 04:55 PM (10 years, 1 month ago) |
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i would for sure be looking into agar, your doing alot of work there for unknow results
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