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fahtster
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Saving bacterial cultures by drying them out 7
#27562340 - 11/29/21 07:54 PM (2 years, 1 month ago) |
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Yoyoyo! First off, let me say that I don’t own a scope so this is going to be based on anecdotal evidence and past experiences and I’m hoping to show youz in the next couple months that this (thread title) is possible. Ive only seen a couple ppl through the years even attempt or work with drying cultures out for whatever reason. Babba yaga being the most recent.. maybe he’ll have some input on this
I dry my cultures out for storage purposes.. I use these lids. They allow me to adjust the GE from 98% covered (during colonization) to 40% exposed (for drying out). I stick them in front of a fan to speed the process up. But I’ve found that drying the cultures out has a great side benefit of cleaning them up from bacteria (if there’s mold, the cultures facked).
I take the summers off so all my cultures get dried up in small 4oz glc grain jars using the lids above^^. I’ve noticed that not only do that cultures bounce back but they bounce back clean and hardy af.
You can also tell how clean a culture is by how fast it dries out.. the grain in a bacterial culture will take twice as long to dry out as clean cultured grains (new drinking game… every time I say culture, you have to drink ).
I recently took an f5 pea+ clone and the inner tissue wasn’t as thick as I would’ve liked and I noticed some ice cream type growth some kernels during recovery after a shake.. the 4oz glc grain jar did finish and was clean enough that I got a quart and 2 more glc jars to colonize but the culture was obviously bacterial. Here’s the 1 qt zipblock of that clone culture:

It’s definitely slightly bacterial.. pins are kind of crapping out after a certain size and I thought it’s a great time to make this post because the two glc jars are showing definite signs of bacteria as well.. first let me say that when I dry the glc jars out, I shake them almost everyday to keep the kernels apart so they dry out faster so the jars you’re about to see look pretty gnarly, but it’s only because they get broken up everyday.
Edit: here’s how that block turned out.. she definitely looks like she’s struggling a bit lol

Here’s one of the f5 glc jars

As you can see, they grow back really thick, are hard to break apart and bruise easily and the kernels stay wet even being in front of a fan constantly. By comparison here’s a known clean culture that was made 9 days after the f5.. barely bruises, breaks apart easily, and is drier even though it’s 9 days older

Edit: here’s the f5 jar 10 days later

So… what I’m hoping to show is that once that f5 culture is completely dried out and let sit for at least a month… (it’ll look something like this:)
 …when I use it to knock up qts, the culture will be clean.
When you see myc dust kind of start to form on the inside of the jar after shaking, it’s probably ready to use
I think it’s a common misconception that all bacteria produce endospores… they don’t. There’s actually a small amount of bacteria that do produce endospores in comparison to how many different bacteria there is.. Obviously, if the culture is infected with an endospore producing bacteria, drying it out isn’t going to help
This going to take a couple months but I’ll keep it updated as things progress
And I should say that there’s no substitute for clean work so that should always be the goal and if you’re seeing bacteria a lot, maybe work on improving your sterile technique… but maybe you have a really sweet culture that is proving difficult to clean on tea agar or something and you don’t want to lose the culture, this could be useful to you
Faht
Edited by fahtster (12/10/21 02:28 AM)
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bigfootscreepyuncl
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Re: Saving bacterial cultures by drying them out [Re: fahtster]
#27562398 - 11/29/21 09:01 PM (2 years, 1 month ago) |
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Goddamn Faht keep it up! How many Nobel Prizes have you won so far?! I'm very interested in seeing how this goes so I can carry cultures over in my off season 
I recently did something similar to this completely by accident. I thought I lost a culture forever after last season but discovered I never cleaned out two of my spawn jars that had some old, very dried out colonized grains in the the bottom. I dropped them onto an agar plate and they took off in about 5 days!

I won't gum up your good thread by asking dumb questions so I'll go look up what glc means 
Thanks for continuing to crank out these awesome TEKs!
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Southerner
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Wow, good shit Faht. Here in the southern US we regularly get dry spells that can last for months. After a nice rain the cubes will be back in full force. I’m thinking the myc also dries out like this in nature. Good catch on the drying killing bacteria as well. Have you ever tried this with other species? I don’t have much experience with anything other than cubes.
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fahtster
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Quote:
bigfootscreepyuncl said: Goddamn Faht keep it up! How many Nobel Prizes have you won so far?! I'm very interested in seeing how this goes so I can carry cultures over in my off season 
I recently did something similar to this completely by accident. I thought I lost a culture forever after last season but discovered I never cleaned out two of my spawn jars that had some old, very dried out colonized grains in the the bottom. I dropped them onto an agar plate and they took off in about 5 days!

I won't gum up your good thread by asking dumb questions so I'll go look up what glc means 
Thanks for continuing to crank out these awesome TEKs!
It’s not THAT big of a deal GLC = grain liquid culture. Check out that link I posted. The drying out culture part 100% works.. I do it all the time.. it’s the knocking back bacteria that I’m hoping to show. I know ppl can clean up cultures with antibacterial agar but I just think it’s really interesting that doing nothing for long enough could do the same thing lol.. if anything, maybe it’ll get some other ppl with scopes interested enough to toy with the idea
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Drboomer
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Re: Saving bacterial cultures by drying them out [Re: fahtster]
#27563298 - 11/30/21 02:51 PM (2 years, 1 month ago) |
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Neat idea.
I've been thinking about giving some lids the Swiss cheese treatment, using a whole sfd and letting the grain dry for culture storage. It would be interesting to see if it kills the bacteria. I know it works with swabs. Letting them for awhile seems to be key.
Definitely going to do it with swabs. Just PC the jar, load with swabs and let them dry in the jar a couple weeks.
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bigfootscreepyuncl
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Re: Saving bacterial cultures by drying them out [Re: Drboomer]
#27567778 - 12/03/21 02:50 PM (2 years, 1 month ago) |
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Couple questions for ya Faht - I'm mostly interested in this for storing cultures during my offseason so I don't have to start from spores when I fire back up, not necessarily saving a bacterial culture.
Is the GLI lid critical to this TEK or could I use polyfill, or would there not be enough GE to allow it to dry?
If your lid is critical do you think I could use the tyvek used as a vapor barrier on the exterior of homes? I have quite a bit of that tyvek laying around and am just trying to avoid sourcing new materials if I don't have to.
As always, thanks for the help Faht, you da' man!
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fahtster
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Yeah the large GE is important because you want it to dry out as fast as possible.. they’ll fruit invitro and shit spores everywhere if it’s not quick enough.. even if you shake em.. they’ll grow off single kernels. The drying out seems to put the myc in stasis where it stays “young” too.. I think the less cell division the better
I haven’t used the home wrap stuff before but I think DuPont makes the tyvek the same way but maybe someone that’s used it will chime in 
Faht
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fahtster
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Re: Saving bacterial cultures by drying them out [Re: fahtster]
#27575494 - 12/10/21 02:11 AM (2 years, 1 month ago) |
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Gettin there!

There’s no longer any condensation even with drastic temp swings which is good. Should dry up faster now that that’s gone
Faht
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bigfootscreepyuncl
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Re: Saving bacterial cultures by drying them out [Re: fahtster]
#27575838 - 12/10/21 09:27 AM (2 years, 1 month ago) |
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How long does it usually take to dry the culture?
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bakedbeings
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Registered: 09/01/20
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good stuff as always faht. this makes so much sense. ive been thinking about giving dry cultures a try for a while and am considering running side by side tests on dry grain spawn vs regular grain spawn thats been sealed and refridgerated. in both cases i would use single grains to start cultures on agar every month or so and see how long they hold up. i say this here just in case youve already done this or something like it and have anything to report
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fahtster
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I have not done that yet but all the PE tubs (except the most recent still growing tub) were all from dried grain from December 2020.
Plz report back whatever your find if/when you try it!
Quote:
bigfootscreepyuncl said: How long does it usually take to dry the culture?
A few weeks where I am now but it rains constantly during the grow season.. when I was in the upper Midwest, it was a couple weeks because of how dry it was
Faht
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Stipe-n Cap


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Re: Saving bacterial cultures by drying them out [Re: fahtster]
#27576258 - 12/10/21 02:40 PM (2 years, 1 month ago) |
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I like to store masters, dedication seems like a cool option. Have you tried this with larger jars yet?
Great post btw.
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fahtster
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Re: Saving bacterial cultures by drying them out [Re: Stipe-n Cap]
#27576298 - 12/10/21 03:08 PM (2 years, 1 month ago) |
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Thanks man.. I think I’ve use 1/2 pint jars (8 oz) really early on but opted for the 4oz because they dry out faster. You really want a jar with a straight jar wall.. a qt jar is a pain in the ass because of that bend at the neck.. water gets caught there and you’d have to use a really long ass needle to get a low degree angle and reach the jar wall so you don’t suck up a bunch of air.
Invitro pins can be a real problem if the grain doesn’t dry out fast enough and the last thing you want in an awesome clone culture/isolate is the introduction of spores
But man.. I really want to see ppl play with this stuff.. it’s been great to me. This culture was dried out for 8 months in a jar in a box

I mean com’n, that’s fackin cool as hell so feel free to try larger amounts/jars whatever and report back
Faht
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Drboomer
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Re: Saving bacterial cultures by drying them out [Re: Stipe-n Cap]
#27576311 - 12/10/21 03:15 PM (2 years, 1 month ago) |
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I like drying spawn to preserve it. I use an sfd on a reg mouth qt jar with an sfd under the ring. If you punch a bunch of extra holes in the lid it dries in a couple weeks. If you aren't careful though some grains will dry before full colonization. This makes a liquid innoculant the best use or atleast a g2g transfer. Sticking it in the fridge after full colonization accelerates drying and keeps invitro pinning down.
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Stipe-n Cap


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Re: Saving bacterial cultures by drying them out [Re: Drboomer]
#27576353 - 12/10/21 03:53 PM (2 years, 1 month ago) |
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This is a great idea, I'm looking forward to trying it out.
I'm a while away from growing yet but when I do this technique is definitely going to be on the list. Fuck storing plates if I can colonize a master and then desicate for storage.
Plates take up less space, but still. I like storing masters of my best clones for quick start up.
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bigfootscreepyuncl
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Re: Saving bacterial cultures by drying them out [Re: Stipe-n Cap]
#27584838 - 12/17/21 12:18 PM (2 years, 1 month ago) |
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Well I'm sure you've seen me mention it in other threads but I've got some bacterial jars trying to do their thang right now. Would I be able to PC some 1/2 pint jars and pour some of my colonized quarts into each jar to start drying them out, like a grain to grain transfer? Grain to jar? lol
I apologize if you already outlined it and I just missed, misread or misunderstood the part of how you get the bacterial grains into the 1/2 pint jars to begin with. I also apologize if that's the dumbest question you read all week lol.
Thanks again Faht!
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fahtster
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If you already have the grains colonized, I’d just pour some in an empty jar and dry those kernels out.. no reason to give the bacteria more food.. know what I mean? Or is that what you’re saying? Yes, that’s totally doable imo
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bigfootscreepyuncl
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Re: Saving bacterial cultures by drying them out [Re: fahtster]
#27584856 - 12/17/21 12:35 PM (2 years, 1 month ago) |
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gotcha! that's what I was trying to ask. I meant PCing empty 1/2 pint jars, not sterilizing 1/2 pints of grain to transfer to. I should have worded that much better but my brain is stuck on stupid today..
Thanks for the help and the timely response, as always! You da man Faht
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fahtster
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So one of the jars (there’s 2) looks pretty dry and I’m curious if ~3 weeks is good enough. Here’s how it looks

I was doing a round of jars and went ahead and made a glc syringe from that ^^^ guy

Knocked up a couple of qts

Normally when the grain aren’t dehydrated I’d leave the GE section of the lid uncovered so the water (the leftover water that can’t be extracted because it’s “stuck” to the kernels) can evaporate and I can spawn the grains and with dehydrated grains like these I’d just toss usually but for these, I covered the GE section and I want to see if bacteria is present to reproduce in the excess water… I’ll check back in a couple days and give the filter a whiff.. this is also how I get a heads up on whether or not the qts are probably going to be bacterial and I can make a decision to either re-pc and re-inoc or try to get colonization and throw it in zipblock if it isn’t a bad infection

Good or bad, I’ll update as things go… I used the Steam pen so if there is bacteria, I’m fairly confident it was already present in the culture and not introduced @ inoculation
I do feel good about it (I guess, obviously or I wouldn’t be trying lol) and from what I’ve researched, bacteria dies within a few hours on a dry surface, but whether or not the rh is low enough or the grains themselves are dry enough is what we will find out! 
Faht
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fahtster
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Re: Saving bacterial cultures by drying them out [Re: fahtster]
#27588787 - 12/20/21 08:29 PM (2 years, 1 month ago) |
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So far, so good.
The grains soaked up the excess liquid and started growing new myc already. Filter smells like mushies. 

Should see visible myc in the qts in 6-7 days.. starting from dry myc adds a couple days time compared to using a hydrated glc jar
Faht
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