|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
AGAR'S GRAIN LC
#21082940 - 01/07/15 03:46 PM (10 years, 12 days ago) |
|
|
I am sharing this b/c RR locked his thread b/c people were bumping it to share. Does RR nor like this LC? I do. I have used this method to expand hundreds of grain jars with ease and speed. Now, with Croncir in change and more fair I will bring his work back to life for the newer member who may have not get to see this amazing method. I am no way braking the rules so dont lock my thread bro. Yes, I did read them
Quote:
agar said:
 Pint jar on right is multispore inoculated (SA). Allowed to colonize near 100%. Syringe is 60 ML w/18 gauge needle. Quart jar on left is sterilized water.
 Aspirated 60 ML sterile water into syringe.
 Injected 60 ML sterile water into colonized grain jar.
 Shook grain jar to liberate mycelium from grain.
 Injected total of 180 ML sterile water into grain jar. (shaking mildly between each injection)
 Aspirated out mycelium laden water from grain jar. (all I could aspirate out of the grain jar was 150 ML) Then, injected mycelium laden water back into sterile water jar.  Approximately 500 ML of Mycelium laden water.  (enough LC for approximately 50....10 ML syringes)
The advantage of this procedure is that you can be assured. The culture in clean. Because you can VISUALLY inspect colonization in the grain jar. (if there is FUNK, you can see it )
That is not the case, with sugar type LC's. (multispore syringe inoculated) They can be contaminated, from day 1. And, you NEVER know it.  Until you inoculate spawn jars with that LC. Then, have every single spawn jar GO SOUTH.
EDIT: I was testing out Agar's LC tek for the first time. As you can see in my post where I am dunking the cakes that their are 12 half-pint, regular mouth PF cakes + the qt cake. So I was able to get 100% success rate with this method on my first try. May not be as fast as other methods but better than MS IMO. Slurry seems to be my new favorite method now though for expanding large amounts of grain spawn in breath taking speeds. I would like to add that I did Agar's tek in open air, taking advantage of the SHIP in the jars. I went on to use his method for at least a hundred other jars with no contams. In open air for any new member who may be reading this. The SHIP's provide an easy way for beginners to inoculate jars and create LC with ease. I know many other members may disagree but I have to question if they have even used SHIP's before and are just trying to promote their own methods and discrediting others without trying them. I can't say I have ever gotten a contamination following this method. Granted, gotta be able to know what a healthy jar looks like which is important to know and this method is actually a good way to see if a questionable jar is actually contaminated. Just make the LC and test out on some agar or other nutrient rich material.

By using this method and taking advantage of the SHIP in the jars I was able to bring my success rate of about 85% of doing open air G2G (bacteria contams suck) to 100% using this LC in open air. I am now a fan of slurry now b/c the colonization times blow my mind and I have some ideas to take advantage of the slurry method as do several other TC's on this site.
Edited by blackdust (01/07/15 05:20 PM)
|
wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,919
|
Re: AGAR'S GRAIN LC [Re: blackdust] 1
#21082951 - 01/07/15 03:48 PM (10 years, 12 days ago) |
|
|
It's just a basic GLC...
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
|
Quote:
wowimflabbergasted said: It's just a basic GLC...
Did you not see that other thread??? This is what he was thinking to do
Quote:
GoldenPreacher said:
So I've been reading up quite a bit, and have constructed a recipe for LC.
95ml H2O from tap 3ml honey 1ml molasses 1 drop Wheat-free soy sauce (I'm a coeliac, aka glutard)
Mix, sterilize 15 min (starting time at 15 psi, leaving air to disperse for 10 min before closing vent), Let cool, inoculate with GT LC, shake, store at 29,5 degr. C until somewhat solid, quartz pebble inside to break up.
That's pretty much it, except for my idea to inject peroxide after sterilization, before injecting GT LC, dunno about concentration yet though.. And also, would adding rice miso instead of or together with soy sauce help?
Advice is greatly appreciated!
|
mushpunx
Fungus Punk


Registered: 04/20/14
Posts: 13,394
Last seen: 2 months, 8 days
|
|
I tried this on a run a long time ago. It worked out ok
--------------------

Amateur Mycologists United
AMU Q&A
|
Yerow
Stranger



Registered: 09/22/14
Posts: 1,206
Last seen: 3 years, 7 months
|
Re: AGAR'S GRAIN LC [Re: mushpunx]
#21083179 - 01/07/15 04:34 PM (10 years, 12 days ago) |
|
|
I might try this out!
|
Fheyr13
Call me, maybe?



Registered: 12/06/14
Posts: 289
Loc: New Mexico
Last seen: 9 years, 2 months
|
|
Seems fairly easy, I'll give it a try. Would it work with partially colonized (50%) jars?
-------------------- If you continue to burn down the herb....
We goin' to burn down the cane fields...
|
wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,919
|
Re: AGAR'S GRAIN LC [Re: Fheyr13]
#21083221 - 01/07/15 04:46 PM (10 years, 12 days ago) |
|
|
I did this for my last PE tubs. I forgot I still have a full 60cc syringe of it in my fridge!
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
Re: AGAR'S GRAIN LC [Re: Fheyr13]
#21083225 - 01/07/15 04:46 PM (10 years, 12 days ago) |
|
|
I was testing out Agar's LC tek for the first time. As you can see in my post where I am dunking the cakes that their are 12 half-pint, regular mouth PF cakes + the qt cake. So I was able to get 100% success rate with this method on my first try. May not be as fast as other methods but better than MS IMO. Slurry seems to be my new favorite method now though for expanding large amounts of grain spawn in breath taking speeds. I would like to add that I did Agar's tek in open air, taking advantage of the SHIP in the jars. I went on to use his method for at least a hundred other jars with no contams. In open air for any new member who may be reading this. The SHIP's provide an easy way for beginners to inoculate jars and create LC with ease. I know many other members may disagree but I have to question if they have even used SHIP's before and are just trying to promote their own methods and discrediting others without trying them. I can't say I have ever gotten a contamination following this method. Granted, gotta be able to know what a healthy jar looks like which is important to know and this method is actually a good way to see if a questionable jar is actually contaminated. Just make the LC and test out on some agar or other nutrient rich material.

By using this method and taking advantage of the SHIP in the jars I was able to bring my success rate of about 85% of doing open air G2G (bacteria contams suck) to 100% using this LC in open air. I am now a fan of slurry now b/c the colonization times blow my mind and I have some ideas to take advantage of the slurry method as do several other TC's on this site.
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
Re: AGAR'S GRAIN LC [Re: Fheyr13]
#21083321 - 01/07/15 05:13 PM (10 years, 12 days ago) |
|
|
Quote:
Fheyr13 said: Seems fairly easy, I'll give it a try. Would it work with partially colonized (50%) jars?
If they are grain jars then yes!
|
jessepinkman
Stranger


Registered: 09/28/14
Posts: 308
|
|
Will definitely give it a rip very soon. Seems less prone to contams than g2g since you dont have to open anything. I just ran into a problem with some g2g.
|
Fheyr13
Call me, maybe?



Registered: 12/06/14
Posts: 289
Loc: New Mexico
Last seen: 9 years, 2 months
|
|
Quote:
blackdust said:
Quote:
Fheyr13 said: Seems fairly easy, I'll give it a try. Would it work with partially colonized (50%) jars?
If they are grain jars then yes!

One of these days I'll figure out how to post GIF and whatnot and I'll be as cool as you guys  Edit: GIFSoup Thanks for all the info, Blackdust.
-------------------- If you continue to burn down the herb....
We goin' to burn down the cane fields...
Edited by Fheyr13 (01/07/15 05:26 PM)
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
Quote:
blackdust said:
Quote:
Fheyr13 said: Seems fairly easy, I'll give it a try. Would it work with partially colonized (50%) jars?
If they are grain jars then yes!

Ummm no. I would wait until the jar was fully colonized. That way even tho you have no way of knowing if the centre is contamed or if there is a risk of endospores that are not visible, at least you will be confident that the culture never stalled and is on the surface healthy.
GLC can work. It does carry some risks IMO but that does not mean someone can't have good success with it. Certain criteria should be met for any grain jar to be considered for a GLC tho.
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
|
Quote:
Pastywhyte said:
. Certain criteria should be met for any grain jar to be considered for a GLC tho.
Yeah! Keep yo bacteria and mold jars behind. 
Your talking to the guy who takes half-colonized PF cakes and slurry them to larger PF bags within 6 days. Contamination? I think not. Others say yes. b/c they are biased against me.

Guys, Whyte is a conservative while I'm more of a liberal. Just to put this into perspective. Whyte is by far a more experienced and knowledgeable cultivator then me though. I look up to him.
|
Mushroom_J
Hard to the Coir !



Registered: 02/17/11
Posts: 774
Loc: East
Last seen: 8 years, 10 months
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21267084 - 02/12/15 11:49 PM (9 years, 10 months ago) |
|
|
If making LC is so risky, how can vendors sell LC culture syringes? (gourmet/medicinal) What's their secret?
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
Making traditional LC is easy if you adhere to proper sterile tek. Their LC's are inoculated with agar wedges and most likely kept and handled with great care. People here are firing spores into sugar water, using tyvek or poly for filters Iinstead of whatmans, this stuff is very risky with liquid culture. Learn agar then do LC. Not only will agar teach you the skills you need to make LC with care, you will also have lots of killer clones ir isolates to really make the most of the speed of LC.
GLC on the other hand has worked for some but, given what I know about endospores I will never risk using it.
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
|
|
Quote:
Pastywhyte said:
GLC on the other hand has worked for some but, given what I know about endospores I will never risk using it.
Honestly, since it's just water, I don't see endospores being more of an issue than with G2G unless the GLC (really LI) is used to inoculate a standard LC.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21267803 - 02/13/15 07:01 AM (9 years, 10 months ago) |
|
|
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.
|
wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,919
|
|
GLC baby, I trust it more than regular LC's, I didn't know it was inferior...?
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
If it works its not inferior. But it has more vectors there is no question. A glc requires your grain master to be 100% clean to be effective. No one can guarantee a grain master is clean visually. There are also extra steps. LC inoculated with a wedge has one vector before aspiration which is inoculation. If you used a wedge you can be very sure its clean. GLC has at least 2 vectors. Inoculation of the grains, and the addtion of the water. Even if the grains are clean it still carries that extra vector. If you then add your myc water to a larger amount of water as outlined above there are even more vectors. Mold is maybe not as big an issue with it but bacteria sure is.
I'm not saying it can't be done I'm saying there is a lot of extra vectors and more risk than many people who advocate it are willing to admit. If you like it then fine. But at least be aware of the pitfalls so you can trace the vector if something goes wrong.
|
wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,919
|
|
|
bw86
Doesn't play well with others


Registered: 11/12/06
Posts: 5,976
Loc: 7b
Last seen: 12 minutes, 44 seconds
|
|
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)
|
FreeWorldOrder


Registered: 12/24/13
Posts: 2,002
Loc: Indiana, USA
Last seen: 11 months, 20 days
|
Re: AGAR'S GRAIN LC [Re: bw86]
#21267996 - 02/13/15 08:26 AM (9 years, 10 months ago) |
|
|
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
Lets Grow Mushrooms Videos
PastyWhyte's Easy Agar TEK
Agar's Liquid Inoculant TEK
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
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.
|
wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,919
|
|
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."
|
FreeWorldOrder


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



Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
|
|
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.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
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.
|
PirateSwazey



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


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
|
|
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.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21269795 - 02/13/15 05:09 PM (9 years, 10 months ago) |
|
|
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.
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
|
|
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.
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,916
Loc: Milky way
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21269974 - 02/13/15 05:48 PM (9 years, 10 months ago) |
|
|
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.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21270001 - 02/13/15 05:53 PM (9 years, 10 months ago) |
|
|
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)
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
|
|
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?
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,916
Loc: Milky way
|
|
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
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21270057 - 02/13/15 06:07 PM (9 years, 10 months ago) |
|
|
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.
|
SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
|
|
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.
-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
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.
|
SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
|
|
I can see that.
-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,916
Loc: Milky way
|
|
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.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
Hmmm that would be interesting. Might be a little more than your average hobbiest would want to do but might have some good applications commercially.
|
SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
|
|
Interesting you mentioned that. eatyualive adds pickling lime to his grains. I asked him a couple of times if that could be one reason he gets away with some of what he does. What do you think?
-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,916
Loc: Milky way
|
|
tetrahydro alpha acids. or tetra iso humulone act as ionophores, very few bacteria have a resistance and are killed by the lack of transport across their membranes.
it acts much better at a lower ph
Edited by Trusted cuItivator (02/13/15 06:37 PM)
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
Its possible. I can't remember his reason for it but that very well might be it.
|
eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
|
|
i recently did 20 jars via GLC and 10 jars via a grain slurry from the same jar. only difference was that i poured the blended up grain into the last 10 jars. the first 20 had liquid mycelia water. slurry jars colonized in 3 days the glc jars took 7-9 and it only took about 15cc per quart jar of grain. i just our directly out of the master into receiving quarts.
the great thing is that one quart jar can inoculate hundreds of jars and you can have full colonization in about 7-9 days. at first it took about 4 days to recover and my slurry jars were 100$% colonized at that time. but on day 4 it began to show recovery. i was about to toss the jars because it didn't do anything for those first few days.
http://www.shroomery.org/forums/showflat.php/Number/21242497#21242497
Edited by eatyualive (02/14/15 01:14 AM)
|
eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
|
|
Quote:
Pastywhyte said: 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.
i find liquids far less work. once you practice it a few times. its like clockwork. in the long run, its doing 1/10th the work. i don't see the logic in it cutting corners. if you know how to prepare a liquid inoculant properly it isn't any more difficult than agar. i find it much easier than agar to be honest. if you can prep a clean culture. pressure cooking some water, blade, jar and using a sterile syringe to inject it isn't that hard. its no more risk than agar. your not adding any nutrients to that equation. and using grains blended in water is about as difficult as making a milk shake and pouring it into a cup. ive used over 5 year old liquid culture syringes from spore works without any issues. i also think it is blended up agar plates in a syringe. its basically the same principle. i even do it with inner stem tissue and distilled water and store it for long periods of time with good success. but it does have a shelf life. apes will last maybe 6 months. other varieties like tex, burma, taz ect will last up to 2 years.
additionally i have been testing out blended grain jars as liquid inoculants. ive been dumping them directly into substrates. out of 6 tubs not one contaminated and ive had several flushes off of each. im working on different spawn ratios to see what the absolute minimum is. but it colonizes the substrate in 3 days!
one of my recent two tub grows involved using 5 quarts of grain. 1 quart was blended with water and poured into the substrate. the other 4 quarts were then spawned normally. the grain, slurry and substrate were all mixed in thoroughly and evenly with gloves on. this colonized in 3 days, i cased the tubs and the tubs looked like this after a week. tub 1
  tub 2
 
Quote:
SpitballJedi said: Interesting you mentioned that. eatyualive adds pickling lime to his grains. I asked him a couple of times if that could be one reason he gets away with some of what he does. What do you think?
old hands used to put jet dry in their grain to use as a releasing agent. it would loosen the gunk off the grain. what i use pickling lime for wasn't intentionally for any type of ph buffering(maybe it helps?). i use dirty ass wbs. so the pickling lime allows the gunk to get off the grain easier. you can also use gypsum. ive used jet dry in the past in soak water as well. if your using cleaner grain it probably isn't necessary. but i use the cheap dirty ass wbs at walmart.
Edited by eatyualive (02/14/15 01:38 AM)
|
d0urd3n
Just call me "D"

Registered: 09/15/10
Posts: 5,237
|
|
Quote:
eatyualive said: ive used over 5 year old liquid culture syringes from spore works without any issues. i also think it is blended up agar plates in a syringe. its basically the same principle.
What makes you think Sporeworks does that? Just curious.
|
eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
|
Re: AGAR'S GRAIN LC [Re: d0urd3n]
#21271588 - 02/14/15 01:40 AM (9 years, 10 months ago) |
|
|
Quote:
d0urd3n said:
Quote:
eatyualive said: ive used over 5 year old liquid culture syringes from spore works without any issues. i also think it is blended up agar plates in a syringe. its basically the same principle.
What makes you think Sporeworks does that? Just curious.
you can see the color of the syringe looks like an agar mix of some sort. it has a golden color like hamloafs grain soak water liquid cultures. im thinking it might be mea. i could be wrong. just a guess.
ive also had one that had a pinkish color to it. all oysters.
|
d0urd3n
Just call me "D"

Registered: 09/15/10
Posts: 5,237
|
|
Interesting. Good to know. Thanks.
|
eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
|
|
something like this color but colonized with mycelia and blended into sterile water. so its a little lighter.

from this thread.
http://www.shroomery.org/forums/showflat.php/Number/20970238/page/1
i really want to try this grain soak water liquid culture but i don't ever soak my grain. i will have to soak some so i can give it a go and break out the old stir plate and stir bars.
also these come in handy.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
|
Quote:
i don't see the logic in it cutting corners. if you know how to prepare a liquid inoculate properly it isn't any more difficult than agar.
Funny enough my concerns have nothing to do with preparation of liquids as they are extremely forgiving in that regard. My main issues are the inoculation methods themselves. When it comes to LC, an agar wedge is 99.99%. I sometimes don't even test (tho I do not recommend that). Spores and tissue are not nearly as reliable and while I am sure many people see great results, its not as good as agar wedges to inoculate. IME bacteria can be present within the centre of a biopsied fruit and I have received enough dirty prints and syringes from lots of sources to never trust spores to 3D sterile media.
I am not saying that they are bad methods. I am saying that they should not be taken lightly. To tell a new cultivator to go ahead and fire a cc of ms solution from his spore syringe into his LC is reckless. To assure someone that just because they don't see anything nasty on the outside of their grain jar means their GLC is guaranteed is misinformed. I have excellent sterile procedure yet I don't trust myself to do some of those things. I cloned a pan cambo fruit once yet even as light and fine a touch that that took, I have seen bacteria pop up on a cube clone more than once. Was I sloppy? Or was some bacteria embedded in that fruit? Given that I can't find anything that definitively says that cubes only grow via turgor after the primordia stage I am inclined to think it was not my technique.
So what does this mean? It means that given your risk tolerance, experience, scale you intend to expand to, and how much time you have, a person should choose their method to fit those criteria. I'm not bashing liquids. I love them. I don't want to see them get a bad rep cause all a new person could find regarding their use was "there great, do it".
|
eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
|
|
Quote:
Pastywhyte said:
Quote:
i don't see the logic in it cutting corners. if you know how to prepare a liquid inoculate properly it isn't any more difficult than agar.
Funny enough my concerns have nothing to do with preparation of liquids as they are extremely forgiving in that regard. My main issues are the inoculation methods themselves. When it comes to LC, an agar wedge is 99.99%. I sometimes don't even test (tho I do not recommend that). Spores and tissue are not nearly as reliable and while I am sure many people see great results, its not as good as agar wedges to inoculate. IME bacteria can be present within the centre of a biopsied fruit and I have received enough dirty prints and syringes from lots of sources to never trust spores to 3D sterile media.
I am not saying that they are bad methods. I am saying that they should not be taken lightly. To tell a new cultivator to go ahead and fire a cc of ms solution from his spore syringe into his LC is reckless. To assure someone that just because they don't see anything nasty on the outside of their grain jar means their GLC is guaranteed is misinformed. I have excellent sterile procedure yet I don't trust myself to do some of those things. I cloned a pan cambo fruit once yet even as light and fine a touch that that took, I have seen bacteria pop up on a cube clone more than once. Was I sloppy? Or was some bacteria embedded in that fruit? Given that I can't find anything that definitively says that cubes only grow via turgor after the primordia stage I am inclined to think it was not my technique.
So what does this mean? It means that given your risk tolerance, experience, scale you intend to expand to, and how much time you have, a person should choose their method to fit those criteria. I'm not bashing liquids. I love them. I don't want to see them get a bad rep cause all a new person could find regarding their use was "there great, do it".
oh i didn't mean using spores. yeah but you could blend up a plate like muda does into distilled water and pour it or inject it with a syringe. yeah exactly pasty. its not for everyone. but it depends on timing really. for me its less work. thats what i like about it most. if i g2g it takes me almost 5 different sittings and clean setups to equal to what i can do in one sitting with 1 1/2 pint pf jar. for instance. if i have a grain master that inoculates 10 quart jars. i would need 5 quart masters to inoculate 50 quart jars. whereas, with liquid, i could easily pressure cook 10 jar batches one day a week so on saturday morning i could do all of the transfer in one shot with 1 pf jar volume of mycelia. whereas, it would take a much higher volume of inoculant and clean sessions to get the same result in half the time it takes for those quarts to colonize using g2g. in the end im doing less work, its convenient, with the same end result. which to me, is extremely helpful when you spend 60 hours a week working.
|
Mushroom_J
Hard to the Coir !



Registered: 02/17/11
Posts: 774
Loc: East
Last seen: 8 years, 10 months
|
|
For safety reasons, couldn't you just PC your LC mix twice, a day apart and also do your PC runs for 1.5 hrs each?
I do my plates for 1 hr 15 minutes at 17 psi just to be safe even though it isn't really needed.
I have well water and it's laden with bacteria. Not sure what the PPM's are but it's hard water. From doing cloning with plants I know the water will grow bacteria and that's just plain water. No nutrients added. I'm guessing it's the oxygen being pumped into it via bubblers. I have to change the water every 3-4 days. My water has a PH of 6.8
Even leaving out a cup of water will grow white slime but not as fast as bubbled water. I'm probably going to die!
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21272027 - 02/14/15 07:09 AM (9 years, 10 months ago) |
|
|
100 G2G jars in a still air room gave me about 80% success rate 100 GLC jars in a still air room gave me about 100% success rate
Thats a lot of jars. GLC worked better for me.
|
SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21272690 - 02/14/15 10:31 AM (9 years, 10 months ago) |
|
|
Part of the reason LC's in general have a bad rep is because of the methods used to make them. There is nothing inherently wrong with LC. But, there may be something wrong with the stuff you put in them.
Having a fast approach is underestimated. A lot of what is commonly done is to provide a much bigger window for colonization to beat out contamination. The bigger the window, the more time we have and the slower we can go. Super fast colonization allows for a smaller window.
I think people run in to issues when people combine some of the fast approach teks with slower approach teks. When this happens, some teks get a bad rap. I think there are some subtitles at work and often get over looked.
-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
|
|
Could you elaborate? I don't understand.
|
SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21273116 - 02/14/15 12:35 PM (9 years, 10 months ago) |
|
|
ummmm, I kinda rather not. It might be too hard to elaborate without sending the wrong message. I just wanted people to take a deeper look at the things they do and criticize.
-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
Edited by SpitballJedi (02/14/15 12:36 PM)
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21273126 - 02/14/15 12:37 PM (9 years, 10 months ago) |
|
|
Yeah, faster methods can be used to beat less sterile media. I have been saying this for a long time. Being clean and sterilizing our media just creates a window for the inoculate to win the race. How big a window we need to colonize depends on how fast we can colonize. I think bacteria is the largest threat in the speed battle.
000 I know this is a piss poor example but how could my oven pasteurize cakes colonize and give fruits if not for the speed of slurry. The 3 day colonization time slurry gave me let me work in a non sterile environment and to use the oven to kill enough enodo spores/bacteria to get fruits.

|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21273167 - 02/14/15 12:50 PM (9 years, 10 months ago) |
|
|
Quote:
blackdust said: 100 G2G jars in a still air room gave me about 80% success rate 100 GLC jars in a still air room gave me about 100% success rate
Thats a lot of jars. GLC worked better for me.
Sorry but that is a ridiculous comparison and your results are predictable. A still air room does not cut the mustard for G2G. Provided your GLC and grain jar lids were equipped with a SHIP, of course your results would be better. The whole point of a SHIP is to allow for use in open air. G2G will never be as successful in a room. That comparison is akin to saying "I drove a quad and never got in an accident. I then drove a car using my feet to steer instead of my hands and got in an accident. Quads are safer than cars."
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
|
Quote:
Pastywhyte said: \ G2G will never be as successful in a room. That comparison is akin to saying "I drove a quad and never got in an accident. I then drove a car using my feet to steer instead of my hands and got in an accident. Quads are safer than cars."
I know and thats why GLC was safer for me b/c I used SHIPS.
These were all done in a still air room. I expanded the spawn by G2G. I would get 1 or 2 bacteria jars for every 10. Always fucking bacteria.
  
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21273251 - 02/14/15 01:12 PM (9 years, 10 months ago) |
|
|
Which is why that's an unfair comparison. G2G should be done in sterile air.
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21273258 - 02/14/15 01:14 PM (9 years, 10 months ago) |
|
|
Quote:
van der griegen said: Which is why that's an unfair comparison. G2G should be done in sterile air.
still makes GLC safer imo. Dont you see all these people with SABS getting bacteria and mold in their tubs.
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 4 years, 1 month
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21273270 - 02/14/15 01:17 PM (9 years, 10 months ago) |
|
|
No, not that many. Usually it's still air bathrooms.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
Re: AGAR'S GRAIN LC [Re: Psilicon]
#21273276 - 02/14/15 01:18 PM (9 years, 10 months ago) |
|
|
Do G2G in a SAB and watch your success rate improve. Its not the fault of the method if you choose to implement it incorrectly. Again I am not bashing GLC but there is a reason commercial operations do not expand with that method. Most use G2G or LI to expand. FrankHorrigan experienced his biggest contam meltdown with GLC. I'm sure most people who are familiar with him agree he knows his stuff. If he can lose 24 tubs with it anyone can. Understand the pitfalls. GLC will work great until it doesn't. The biggest risk with liquids especially GLC is that when it does let you down, it lets you down big.
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
|
Considering that 1 master jar of GLC can produce 100x as much as 1 G2G master jar and I can test out a GLC and know if its good or not within a week is the reason it has my love. Not to mention that I dont have to work in a little plastic box. Testing out a GLC before MASSIVE expansion is like following up with a MASSIVE institution ~ Gotta do it or deal with the risk of getten fucked.
Is this not what agar is for? Talk about it enough. lol
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21273339 - 02/14/15 01:38 PM (9 years, 10 months ago) |
|
|
That is what agar is for. If your inoculating your master with agar and testing it with agar then you have little to worry about. But if I was using agar I would just inoculate LC and remove the extra stick of the SHIP or the risk of germinated endospores. Takes less time to colonize an LC with a nice agar wedge than it does to colonize a grain master, if speed is your concern.
|
SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21273341 - 02/14/15 01:39 PM (9 years, 10 months ago) |
|
|
Quote:
blackdust said: I can test out a GLC and know if its good or not within a week is the reason it has my love.
Are you saying that you can take a glc, inoculate more jars, spawn those jars, and get fruits within a week?
Or are you saying you test it out on agar for contams first?
Considering that PCing doesn't kill everything on grains, I hope you test that GLC regularly if you store it.
-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
Edited by SpitballJedi (02/14/15 01:41 PM)
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
|
Quote:
SpitballJedi said:
Or are you saying you test it out on agar for contams first?
Considering that PCing doesn't kill everything on grains, I hope you test that GLC regularly if you store it.
Yes. I actually tested my fist GLC on cakes as shown in the OP. I would recommend testing on agar before using to not repeat what happened to Frank. Or test on a small batch of cakes and then slurry the cakes to grain jars. 1 cake can make 50 grain jars. I want to try this GLC again. Now that I know Eat can fully colonize a PF cake in 4 days I want to try the same with a GLC instead of steam tissue.
|
Yerow
Stranger



Registered: 09/22/14
Posts: 1,206
Last seen: 3 years, 7 months
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21273492 - 02/14/15 02:25 PM (9 years, 10 months ago) |
|
|
Think it would help/do anything to let that water sit in the jar for a few hours, maybe shake a few times in that time, to really get that myc out into it?
|
eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
|
|
back from the dead. good to see you bd.
hey sbj been a long time man how ya been?
oh guess what i just made sbj?
i made all my tub hole magnets! colonizing 4 tubs now with the magnets on and then ill put the Dilated filters on next during fruiting. i did 4 tubs with 8 quarts of spawn to 11 quarts of substrate. sub depth was about 4". going to case in about 3 days when the subs finish colonizing. i haven't been able to beat my 3 day bulk colonization average yet. but im trying hard. i don't think it can get any faster. i did make some slight modifications. i would usually spawn my tubs with the polyfil in the holes and put the tub in a draft free area. now ive started using the magnets to colonize the tubs in my grow room with the fan moving air. then switch up to poly and dilated filters when fruiting. the issues i was having for the last several months i have worked out. which was directly related to how tight i was stuffing my bottom polyfil holes. i had them in, but not solid bulging tight. i made that minor change, built a new grow setup with a small fan and its working rock solid now. i also started using a 6500 light rather than natural daylight. the ceiling fan wasn't cooperating lol. the lighting isn't making much of a difference other than the fruits grow directly toward it rather than all over the tub with diffused light. yields still the same in cased and uncased bulks, natural diffused light or artificial light. doing about 6-9z dry average per tub. ill pm ya the magnet pics.
Quote:
SpitballJedi said:
Are you saying that you can take a glc, inoculate more jars, spawn those jars, and get fruits within a week?
GLC takes me about 7-9 days for full colonization in quart grain jars. for grain slurry to bulk=2 weeks from inoculation to harvest. 12-13 days on average for slurry. but a GLC takes me about 5 extra days to colonize quarts. so for GLC 18 days till fruit from inoculation of grain jars to harvest when your spawning to bulk tubs. if you fruit directly from bottles that would decrease the time by a few days up to a week.
i don't use a SAB for g2g or any of that and i get near 100% success without contamination. i don't think the method matters. if its a flowhoood, shmuv, sab, its your technique and following the principle using that method that ensures success. ive used all 3 and the contam rate is the same.
|
SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
|
|
Hey eat, good to see you. It has been a while. Glad to see you're still kicking ass. 
The time scale you mention seems reasonable.
Like PW mentioned, why make GLC instead of Agar LC? We've talked enough to know this is an honest question. You've inspired me to question my methods in the past and I've actually changed some things to make it easier because of it.
-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
|
eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
|
|
Quote:
SpitballJedi said: Hey eat, good to see you. It has been a while. Glad to see you're still kicking ass. 
The time scale you mention seems reasonable.
Like PW mentioned, why make GLC instead of Agar LC? We've talked enough to know this is an honest question. You've inspired me to question my methods in the past and I've actually changed some things to make it easier because of it.
your not really making a GLC. its almost already there before you. you simply blend up the grain in the water and you have a very large volume of LI for inoculation. i guess you could say for convenience. ill give an example, i g2ged 10 quart jars. thats all i have going(maybe you have a situation where your growth stalled or life got in the way). i want to spawn 2 tubs and need a quick turnaround to get more spawn going for another round of tub spawning and flushes. i could 1)g2g one last quart and make 10 more quarts for spawn. or i could keep that 1 grain jar and make enough inoculant for over 100 quart jars. its easy because you already have a large volume of culture that you can transfer. you could use a plate. but it all really depends on the situation. if your using an LI to inoculate a bunch of grain jars, use that last grain jar for a GLC. and then you spawn the other 9 jars. then that one GLC turned into enough spawn for 20 tubs or more.
1 its easy. and if you dont' have any agar plates or anything of that nature you can use your last remaining grain jar as a GLC to transfer and make a ton more jars.
2. that one grain jar can inoculate enough jars comparable to 10 or more plates of LI. you could also use a grain from that remaining quart and put it on agar. but then you would need to grow that out on a plate. when you already have a large volume of inoculum at your disposal. the way i look at it is, if you hav a quart volume of liquid inoculant. you have an unlimited amount of spawn that you can make in a moments notice, and its convenient and not that difficult.
Edited by eatyualive (02/14/15 09:28 PM)
|
Neomorph



Registered: 12/10/14
Posts: 767
Loc: Europe
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21278349 - 02/15/15 03:17 PM (9 years, 10 months ago) |
|
|
Nice work dude, the jar has only water or it has honey to ?
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
Re: AGAR'S GRAIN LC [Re: Neomorph]
#21278352 - 02/15/15 03:18 PM (9 years, 10 months ago) |
|
|
Quote:
kastriotr said: Nice work dude, the jar has only water or it has honey to ?
distilled water that has been pressure cooked along with the syringe.
|
blazedup
Dirk Diggler


Registered: 06/05/11
Posts: 1,240
Loc: Middle Earth
Last seen: 4 months, 15 days
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21313151 - 02/22/15 01:40 PM (9 years, 10 months ago) |
|
|
After completing this do you toss the grain in the jar or will it recover? I am about to try this out.
|
blackdust


Registered: 02/28/09
Posts: 8,327
|
Re: AGAR'S GRAIN LC [Re: blazedup]
#21313248 - 02/22/15 01:48 PM (9 years, 10 months ago) |
|
|
let the jar recover.
|
blazedup
Dirk Diggler


Registered: 06/05/11
Posts: 1,240
Loc: Middle Earth
Last seen: 4 months, 15 days
|
Re: AGAR'S GRAIN LC [Re: blackdust]
#21313275 - 02/22/15 01:54 PM (9 years, 10 months ago) |
|
|
Quote:
blackdust said: let the jar recover.
Thanks
|
TrIpPyDuDe
FUCK


Registered: 12/07/14
Posts: 1,263
Loc: Kentucky
Last seen: 9 years, 9 months
|
Re: AGAR'S GRAIN LC [Re: blazedup]
#21313311 - 02/22/15 02:03 PM (9 years, 10 months ago) |
|
|
I just bought pre poured agar plates. Can't wait to use them
|
|