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invitro


Registered: 05/03/13
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Speed and effectiveness of liquid cultures
#19326177 - 12/25/13 04:46 PM (10 years, 1 month ago) |
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I have a few thoughts on making LC's Some might see this post as nit-picky but I want to have the best LC I can possibly have, so:
When Blue Helix said that stirring the LC constantly increases how quickly a lc finishes, is this because of increased oxygen in the fluid? My guess is that having all the blobs of myc bump into eachother hinders growth somewhat significantly. But maybe if they break off they can start new colonies.
http://www.shroomery.org/forums/showflat.php/Number/5484818
If oxygen is the key perhaps there are ways to oxygenate the fluid without stirring. I stopped swishing one of the lc jars and I can see the individual blobs are growing, I don't want to swish them any more than I have because it'll smash all those outer hypae.
---- Next thought, if you have a lc that is say 5% colonized with mycelium and you have one that is 50% colonized, does it make a difference when it comes time to inoculate a grain jar?
I also don't really understand the need for breaking up the mycelium so that you can suck it up into a syringe. If it gets stuck, just push it out and suck it back in again. This always clears it up for me.
Edited by invitro (12/25/13 04:53 PM)
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cronicr



Registered: 08/07/11
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Re: Speed and effectiveness of liquid cultures [Re: invitro]
#19326233 - 12/25/13 05:02 PM (10 years, 1 month ago) |
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That's part of the reason to stir and break up but also to move the nutes around or they settle where as myc will try to float. As for clogging get a better gauge
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newera
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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19326388 - 12/25/13 06:06 PM (10 years, 1 month ago) |
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You also wanna break it up before drawing to spread the myc when u inoculate
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invitro


Registered: 05/03/13
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Re: Speed and effectiveness of liquid cultures [Re: newera]
#19326481 - 12/25/13 06:40 PM (10 years, 1 month ago) |
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I can see doing that at the end, usually it gets pretty well broken up as I suck it into the syringe.
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invitro


Registered: 05/03/13
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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19329361 - 12/26/13 03:36 PM (10 years, 1 month ago) |
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Cronicr, What gauge do you recommend for thick lcs? 16g seems to be ok.
How long would you pc a 60cc syringe full of fluid at 15 psi?
Also, how come nobody uses coffee as part of the lc mix? I'm getting ready to pc some coffee + other nutrients into an old honey-water lc to see if it will resume growth.
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cronicr



Registered: 08/07/11
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Re: Speed and effectiveness of liquid cultures [Re: invitro]
#19329382 - 12/26/13 03:41 PM (10 years, 1 month ago) |
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no coffee because they only get a short pc time and it's better to saok your grains in , the lc's nutes are limited and should be and i use 18 gauge but it's not absolutely necessary just handy, and a 60 cc syringe should be fine to chuck in with your lc and get the same time
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invitro


Registered: 05/03/13
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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19329423 - 12/26/13 03:57 PM (10 years, 1 month ago) |
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I don't understand the part about coffee only getting short pc time? I have been soaking grain in coffee though, but thought if it's good for grains it must be good for lc.
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cronicr



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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19329437 - 12/26/13 04:01 PM (10 years, 1 month ago) |
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hers wha i did, not that i reccomend it because of the obviuos reasons, i started a brf/water mix and sterilized it in an old spice jar
 i let it colonize enough to transefer to another one
 i let that colonize
 and took a 20 cc syringe of sterile water and squirted it in, closed the lid tight and shook it up, then sucked up the water back up and inoculated my jar/jars
 not much of a hassle and i could have done 20 jars and g2g'd those to ten more each if i wanted
you can give the coffee a shot i just don't see the need to risk it
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36fuckin5
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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19329446 - 12/26/13 04:06 PM (10 years, 1 month ago) |
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Using coffee means you have to balance your pH. Coffee is very acidic. Lime and gypsum do this in grain/bulk, but with your suggestion, you won't have anything balancing it.
Just skip it. I promise it won't be worth the hassle.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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invitro


Registered: 05/03/13
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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19329456 - 12/26/13 04:09 PM (10 years, 1 month ago) |
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It's almost pure curiosity, but there is also the chance of coming up with something way more effective. Thanks for the write up about the brf water. According to violets write up on brf water she separates the milky water from the "clear" water, or less milky water. You water seems to be very milky, is that straight grain water? Also it looks very congealed, what is the consistency there?
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cronicr



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Re: Speed and effectiveness of liquid cultures [Re: invitro]
#19329474 - 12/26/13 04:12 PM (10 years, 1 month ago) |
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there's no water in those pics lol, yes it will sorta be milky but i never had any issues with anything i just did it for the fuck of it with no real hope but it worked great , i'm trying it with actual agar this week for my panaeolus grow
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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


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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19334098 - 12/27/13 06:54 PM (10 years, 1 month ago) |
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Well I whipped up a jar just like the one you showed, I had the rubber lid facing down, also a ship and a 3/4 inch sfd held in with rtv. I got a little condensation on the surface after I took it out of the pc (maybe .5 cc of water?), plus the liquid of the syringe made it a little to wet for me (so I tilted it sideways and hopefully some spores remain on the dryer agar). How do I get rid of the moisture? What is the point of going rubber up, besides the fact that it might be slightly easier to remove the lid? I was pretty skimpy on the agar too, it was pretty soft, I didn't measure it out really just guestimated.
I'd like to get it super dry like your looks, any help would be appreciated.
edit: I just recalled that I forgot to put foil over the lid when pcing, maybe that was the problem.
Edit: I did 3 more 1/2 pint jars of rice water agar, each with 1.5% agar, one with rubber down, two with rubber up. They all ended up pretty good, not a drop of moisture on surface, but small drops on the walls of the jar. I used tin foil this time. I would still like to know why anyone would go rubber up on the lid if they have a sfd.
Edited by invitro (12/28/13 12:22 AM)
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Icyus
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Re: Speed and effectiveness of liquid cultures [Re: invitro]
#19334107 - 12/27/13 06:59 PM (10 years, 1 month ago) |
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Add oxygen to water... would algae work?no expert in mycologi, so it would be a wild guess, but a guess just the same...
-------------------- And thus begins the reverse-fusing of our one-dimentional understanding, and adds ever-expanding perspectives, in depth and number; splitting our perception, and in so doing, seemingly irrationally, creates yet more one-ness, with all that ever was, is and will ever be, streching across the infinite, inunderstood concept of everything, percievable and not.
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Quexl

Registered: 12/17/13
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Re: Speed and effectiveness of liquid cultures [Re: Icyus]
#19334288 - 12/27/13 08:02 PM (10 years, 1 month ago) |
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Yeah, LCs need to be CLEAN CLEAN CLEAN is my understanding. but I LOVE the idea of synergistic systems, so hers my 2 cents:
Many strains of algae actually behave like bacteria, not plants (cyanobacteria aka 'green algae' for example). That would be a bad competitor for nutrients and oxygen for what you want to grow, cyanobacteria would crash every wanted parameter to 0 (DO, nutrient conc.). Bacterial algae, unless their benthic, suck oxygen like crazy and produce dissolved CO2, carbonic acid, and a whole range of nasty stuff for a LC.
Benthic nitrogen-fixing or anaerobic bacterial algae (biofilm) basically produce ammonia and methane. So no dice there. 
aaand.
Surface area of a fluid = increased oxygen interface. That's why people swirl brandy to get a good whiff of it. I've tried LCs (honey) 4 times without polyfill gas exchange using laboratory equipment (mag-stir and incubation) and always came up with bunk. All I got were polymer filaments from the honey. At least at 1/2 pint size, gas exchange seems to be required, negating the use of other organisms.
If you can get it to work, record everything.
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kaptanoblivious
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Re: Speed and effectiveness of liquid cultures [Re: Quexl]
#19334342 - 12/27/13 08:16 PM (10 years, 1 month ago) |
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Am nothing but a noob, but I recently did two LCs with no gas exchange at all, and they were very successful. got about a pint of thick LC after one week at ~80C, just shaking the jar twice per day. I used corn syrup supplemented with a bit of oatmeal supernatant. Used them to innoc some BRF cakes, and no contams yet in 12 jars, 80% colonized.
http://www.shroomery.org/forums/showflat.php/Number/19304576
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Quexl

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Re: Speed and effectiveness of liquid cultures [Re: kaptanoblivious]
#19334365 - 12/27/13 08:21 PM (10 years, 1 month ago) |
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Awesome job Kap!,
Were those 1 pint jars? Did you use gasket seals w/o gas exchange with stirring? That's how I want my LCs to turn out! Time for me to switch nutrient mediums I think.  anyway, off topic. PM me.
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cronicr



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Re: Speed and effectiveness of liquid cultures [Re: invitro]
#19336883 - 12/28/13 12:29 PM (10 years, 1 month ago) |
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Quote:
invitro said: Well I whipped up a jar just like the one you showed, I had the rubber lid facing down, also a ship and a 3/4 inch sfd held in with rtv. I got a little condensation on the surface after I took it out of the pc (maybe .5 cc of water?), plus the liquid of the syringe made it a little to wet for me (so I tilted it sideways and hopefully some spores remain on the dryer agar). How do I get rid of the moisture? What is the point of going rubber up, besides the fact that it might be slightly easier to remove the lid? I was pretty skimpy on the agar too, it was pretty soft, I didn't measure it out really just guestimated.
I'd like to get it super dry like your looks, any help would be appreciated.
edit: I just recalled that I forgot to put foil over the lid when pcing, maybe that was the problem.
Edit: I did 3 more 1/2 pint jars of rice water agar, each with 1.5% agar, one with rubber down, two with rubber up. They all ended up pretty good, not a drop of moisture on surface, but small drops on the walls of the jar. I used tin foil this time. I would still like to know why anyone would go rubber up on the lid if they have a sfd.
for condensation i always wait till my stuff is back to room temp before i put the lid on and toss in the pc, and rubber is really only for the reason you stated, i make my brf/water mix much drier/thicker then the agar substitute tek calls for, i also don't use foil on much these days but the moisture /condensation should go away over the course of a few days anyway so it's not a big deal
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Icyus
KavitārkikasiṃHa



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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19336941 - 12/28/13 12:45 PM (10 years, 1 month ago) |
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Seriously... would algae work? I thought it a good idea... if it doesnt kill the spongebobs..
-------------------- And thus begins the reverse-fusing of our one-dimentional understanding, and adds ever-expanding perspectives, in depth and number; splitting our perception, and in so doing, seemingly irrationally, creates yet more one-ness, with all that ever was, is and will ever be, streching across the infinite, inunderstood concept of everything, percievable and not.
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invitro


Registered: 05/03/13
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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19336950 - 12/28/13 12:47 PM (10 years, 1 month ago) |
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Why do you say you don't recommend what you did for obvious reasons? The more I think about it the more I like that technique. Just out of curiosity, why is there a depression in your agar, the center seems lower than the sides. The ones I do turn out perfectly flat.
Also your agar is pretty white, not that it matters but if you take the rice soak water and let it settle and then skim the top liquid, it's nearly clear. You can look up through the bottom of the jar and see what's on top, esp if you keep your agar patty thin.
The problem with algae is, how would you get the contams off the algae? It's probably a lot more trouble than it's worth but if you feel the need to try it go for it.
Edited by invitro (12/28/13 12:49 PM)
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cronicr



Registered: 08/07/11
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Re: Speed and effectiveness of liquid cultures [Re: invitro]
#19336971 - 12/28/13 12:51 PM (10 years, 1 month ago) |
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Quote:
invitro said: Why do you say you don't recommend what you did for obvious reasons?
i just wasn't as sterile as i should have been lol! i was never really expecting much and the reason the color is off and there are depressions is because it isn't agar at all it's just brf/water
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invitro


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Re: Speed and effectiveness of liquid cultures [Re: cronicr]
#19337016 - 12/28/13 01:05 PM (10 years, 1 month ago) |
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I missed the part about brf, I though you meant brf soak water.
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cronicr



Registered: 08/07/11
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Re: Speed and effectiveness of liquid cultures [Re: invitro]
#19337017 - 12/28/13 01:06 PM (10 years, 1 month ago) |
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the same thing for agar though i just didn't have a pc on hand when i did it
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