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bw86
Doesn't play well with others


Registered: 11/12/06
Posts: 5,976
Loc: 7b
Last seen: 12 minutes, 29 seconds
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I hate all GLC LI or LC. i sipmly dont use them or have a reason to.
G2G/ agar or go home. playing wih all that jazz is unneeded. maybe there is higher risk of contamination.But not if you just do everything right.
Edited by bw86 (02/13/15 08:14 AM)
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FreeWorldOrder


Registered: 12/24/13
Posts: 2,002
Loc: Indiana, USA
Last seen: 11 months, 20 days
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Re: AGAR'S GRAIN LC [Re: bw86]
#21267996 - 02/13/15 08:26 AM (9 years, 10 months ago) |
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I've inoculated dozens of jars from a master and have yet to have a single jar or grow contaminate due to using this method. Not one.
Especially when the receiving jar has a SHIP.
Nothing wrong with g2g either as long as it is done right...
-------------------- "They who can give up essential liberty, to obtain a little temporary safety, deserve neither liberty nor safety." - Benjamin Franklin
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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I did 500+ jars with G2G no issues. Then I got a batch of grain with a high endospore count. Ended up losing a lot before I figured it out. With GLC it would have been worse.Moral is that ever method has its downside and that blind faith in a method is foolish. Become fluid in your approach, learn about the drawbacks to every method and you can't be beat. My problem is not with any method in particular. Its with people making claims of infalliblity and refusing to admit to potential vectors.
The best thing anyone can do is learn proper sterile tek. Agar will teach that. Then no matter how yiu expand you have the ability to overcome any issue.
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wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,919
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Totally Pasty. I admit any possible contam vectors and am not advocating one method over another. I use a lot of methods for inoculation. LC, GLC, LI, G2G, agar, spore syringes...and I love what you said up above. "If it works its not inferior."
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FreeWorldOrder


Registered: 12/24/13
Posts: 2,002
Loc: Indiana, USA
Last seen: 11 months, 20 days
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Well said brother...
-------------------- "They who can give up essential liberty, to obtain a little temporary safety, deserve neither liberty nor safety." - Benjamin Franklin
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PastyWhyte's Easy Agar TEK
Agar's Liquid Inoculant TEK
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PirateSwazey



Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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I heard even Aloha's most common procedure is agar --> G1 --> G2 spawn bags, and they are the ''largest producer of american grown medicinal mushrooms'' in the world. Shouldn't bee too much work for a small-time home cultivator to keep up with just a few plates and jars lol. To each his own... Sometimes I play with liquid cultures & inoculants but I'd always prefer to not.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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I completely respect that. LC is the big payoff that carries a big risk with it. The level of care that needs to be exercised for consistently good results is not something that many people are willing to do. People who see LC or any liquid inoculate as a way to cut corners are just setting themselves up for a nasty lesson. It might not be this grow or the next, but it will come. Depending on how complacent they are when it does come it can really hurt.
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PirateSwazey



Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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If you really enjoy this as a hobby then it shouldn't be much like work (culturing on agar) anyways
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Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
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Quote:
Pastywhyte said: Germinated endospores in a glc will spread out more in the liquid allowing a far greater spread of inoculation points than with a simple G2G. Given that bacteria recovers immediately this would give them a significant advantage over your cube mycelium.
I don't follow. I can't see how it could spread any farther than a properly shaken G2G recipient, which sees an even distribution of grains throughout the jar.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: AGAR'S GRAIN LC [Re: Psilicon]
#21269795 - 02/13/15 05:09 PM (9 years, 10 months ago) |
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Bacteria blooms in water. A single grain with bacteria can easily compromise an entire quart of water and will travel fast through it as well. While I have no doubt that that same single contamed grain would spell trouble in grains you can't tell me that you think the single act of shaking the jar would compromise all the grains in it 
Often when bacterial endospores are an issue its only with a very small number of the total grains in a jar. There is no way that they are going to comprise nearly as many grains through shaking as they would in a liquid suspension. Chances are that if you had a single grain with bacteria in a master jar that you G2G you might have a small issue with 1 or 2 of the other jars (maybe not even that much as often the bacteria is in the centre of the grain and will not spread quickly if the grain has already been colonized). Adding the water to it, then aspirating the water and then expanding it to more water is going to really increase how much ground it can cover.
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Psilicon
Really Nice Guy


Registered: 08/26/12
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Last seen: 4 years, 1 month
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Quote:
Pastywhyte said: Bacteria blooms in water.
It does not! I've cultivated a lot of bacteria, both intentionally and unintentionally, in a lab and in my home. It only blooms in places with available nutrients, and a wash over a grain jar that's undergone total or near-total bioconversion isn't one of those.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,916
Loc: Milky way
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Re: AGAR'S GRAIN LC [Re: Psilicon]
#21269974 - 02/13/15 05:48 PM (9 years, 10 months ago) |
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I can concur with that. biofilm appears if nutrients are avail. in tap water or any other non-nutrient water bacteria kind of just hangs out. actually in my city the PPM of chlorine and clorine compounds are so high the water itself is a low level sanitizer. it reduces contamination between 4 and 5 log.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: AGAR'S GRAIN LC [Re: Psilicon]
#21270001 - 02/13/15 05:53 PM (9 years, 10 months ago) |
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Okay then it certainly spreads easier. If a grain centre is contamed with a small amout of germinated endospores say, 250000, they are gonna disperse through the act of injecting 60 cc if water into their vicinity a lot more efficiently than they are just by having the grain that they are in make contact with other grains in the master. Water has this tendency to wick bacteria. I would also hazard that some nutrients are going to ride along with the myc when you aspirate the water back into the syringe. Sure maybe not much but more than 0.
Edit: in fact if I recall Franks agar media log correctly he made agar using water he dunked his grains in as the sole media source and had lots of growth.
Edited by Pastywhyte (02/13/15 05:59 PM)
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Psilicon
Really Nice Guy


Registered: 08/26/12
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Last seen: 4 years, 1 month
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Quote:
Pastywhyte said: Water has this tendency to wick bacteria. I would also hazard that some nutrients are going to ride along with the myc when you aspirate the water back into the syringe. Sure maybe not much but more than 0.
If you use your GLC immediately, as most recommend, the nutrients don't matter much at all because it obviously can't spread in no time flat. It miiight wick bacteria. I've never heard of that, though. Is it something you've experienced?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,916
Loc: Milky way
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we should think of endospores in CFU, colony forming units. I have access to cyclohexamide agar. it kills all eukaryotes. send me some high endospore grains I can plate them before and after sterilization and see which prokaryotes survive, all colonizes you can see on the differential agar are assumed to grow up from a single bacterial endospore or bacterium hence CFU. it would be interesting to see what is actually growing too, you could do the catalyse test, and determine if you're gram positive or gram negative, and if you have cocci rods or spirals. I would assume they're lactic or acetic acid gram positive catalase positive facultative anareorbes. there's actually things you could treat your grains with to prevent their contamination before even sterilizing, like iso-humulone
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: AGAR'S GRAIN LC [Re: Psilicon]
#21270057 - 02/13/15 06:07 PM (9 years, 10 months ago) |
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If there are nutrients present it will feed on them. The fact that your injecting them into a grain jar of nutrients means it doesn't even matter if it blooms or not. As for the wicking if you doubt that water does not act as a medium they can travel through with ease then the notion that pooling wet grain jars favor bacteria is thus invalidated. So go ahead, ensure that your grain jars have pooling water in them. If your take is correct then bacteria should not be any issue.
Great idea bodhi. Good luck convincing people who think agar is too much of a pain and can't be bothered to inoculate a LC with a wedge and instead would make a grain master and expand with GLC to do that.
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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I rarely quote RR, but he's said that GLC has a lot of bacteria in it that can be seen under a microscope.
It seems to me that the weekend state of mycelium is a perfect opportunity for these bacteria to multiply, in part because the water is very nutrient rich.
I think this follows the same logic as to why LC from MS can be unreliable. The difference is that GLC at least already has some myc in it, but I don't think it recovers as fast as bacteria.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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I have put endospore contamed grains on plates (ones where the bacteria was not immediatly apparent in the jar) and by the time the myc had fuzzed up enough to leap off, bacteria had already run rampant all over the plate. The recovery time for bacteria is immediate for our purposes.
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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I can see that.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,916
Loc: Milky way
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It would be interesting to make hopped grains and adjust the ph to around 4 or under, if cube myc can tolerate it. I bet your bacteria problems would be moot forever.
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