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Invisibleblackdust
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Registered: 02/28/09
Posts: 8,327
AGAR'S GRAIN LC
    #21082940 - 01/07/15 03:46 PM (9 years, 22 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. :rockon:
(enough LC for approximately 50....10 ML syringes)

The advantage of this procedure is that you can be assured.
The culture in clean.:grin:
Because you can VISUALLY inspect colonization in the grain jar.
(if there is FUNK, you can see it :rolleyes:)

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. :frown:
Until you inoculate spawn jars with that LC.
Then, have every single spawn jar GO SOUTH. :eek: :mad2:








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)


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Invisiblewowimflabbergasted
supercalifragilistic
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Registered: 07/16/12
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Re: AGAR'S GRAIN LC [Re: blackdust] * 1
    #21082951 - 01/07/15 03:48 PM (9 years, 22 days ago)

It's just a basic GLC...


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Invisibleblackdust
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Posts: 8,327
Re: AGAR'S GRAIN LC [Re: wowimflabbergasted]
    #21083125 - 01/07/15 04:19 PM (9 years, 22 days ago)

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!





:facepalm:


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Offlinemushpunx
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Registered: 04/20/14
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Re: AGAR'S GRAIN LC [Re: blackdust]
    #21083159 - 01/07/15 04:28 PM (9 years, 22 days ago)

I tried this on a run a long time ago. It worked out ok


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OfflineYerow
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Re: AGAR'S GRAIN LC [Re: mushpunx]
    #21083179 - 01/07/15 04:34 PM (9 years, 22 days ago)

:raisemyglass: I might try this out!


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OfflineFheyr13
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Re: AGAR'S GRAIN LC [Re: blackdust]
    #21083181 - 01/07/15 04:34 PM (9 years, 22 days ago)

Seems fairly easy, I'll give it a try. Would it work with partially colonized (50%) jars?


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Invisiblewowimflabbergasted
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Re: AGAR'S GRAIN LC [Re: Fheyr13]
    #21083221 - 01/07/15 04:46 PM (9 years, 22 days ago)

I did this for my last PE tubs. I forgot I still have a full 60cc syringe of it in my fridge!


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Invisibleblackdust
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Registered: 02/28/09
Posts: 8,327
Re: AGAR'S GRAIN LC [Re: Fheyr13]
    #21083225 - 01/07/15 04:46 PM (9 years, 22 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.


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Invisibleblackdust
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Registered: 02/28/09
Posts: 8,327
Re: AGAR'S GRAIN LC [Re: Fheyr13]
    #21083321 - 01/07/15 05:13 PM (9 years, 22 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!

:awethumb:


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Invisiblejessepinkman
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Registered: 09/28/14
Posts: 308
Re: AGAR'S GRAIN LC [Re: blackdust]
    #21083332 - 01/07/15 05:16 PM (9 years, 22 days ago)

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.


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OfflineFheyr13
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Re: AGAR'S GRAIN LC [Re: blackdust]
    #21083336 - 01/07/15 05:18 PM (9 years, 22 days ago)

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!

:awethumb:




One of these days I'll figure out how to post GIF and whatnot and I'll be as cool as you guys :wink:
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)


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InvisiblePastywhyteMDiscord
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Registered: 09/15/12
Posts: 37,808
Loc: Canada Flag
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Re: AGAR'S GRAIN LC [Re: blackdust]
    #21131403 - 01/16/15 09:56 PM (9 years, 13 days ago)

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!

:awethumb:




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.


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Invisibleblackdust
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Posts: 8,327
Re: AGAR'S GRAIN LC [Re: Pastywhyte]
    #21132559 - 01/17/15 07:19 AM (9 years, 12 days ago)

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. :rofl:


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. :rofl:




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.


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OfflineMushroom_J
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Re: AGAR'S GRAIN LC [Re: blackdust]
    #21267084 - 02/12/15 11:49 PM (8 years, 11 months ago)

If making LC is so risky, how can vendors sell LC culture syringes? (gourmet/medicinal)
What's their secret?


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InvisiblePastywhyteMDiscord
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Re: AGAR'S GRAIN LC [Re: Mushroom_J]
    #21267574 - 02/13/15 04:45 AM (8 years, 11 months ago)

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.


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OfflinePsilicon
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Re: AGAR'S GRAIN LC [Re: Pastywhyte]
    #21267734 - 02/13/15 06:30 AM (8 years, 11 months ago)

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.


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InvisiblePastywhyteMDiscord
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Re: AGAR'S GRAIN LC [Re: Psilicon]
    #21267803 - 02/13/15 07:01 AM (8 years, 11 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.


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Invisiblewowimflabbergasted
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Re: AGAR'S GRAIN LC [Re: Pastywhyte]
    #21267876 - 02/13/15 07:49 AM (8 years, 11 months ago)

GLC baby, I trust it more than regular LC's, I didn't know it was inferior...?



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InvisiblePastywhyteMDiscord
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Re: AGAR'S GRAIN LC [Re: wowimflabbergasted]
    #21267940 - 02/13/15 08:07 AM (8 years, 11 months ago)

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.


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Invisiblewowimflabbergasted
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Re: AGAR'S GRAIN LC [Re: Pastywhyte]
    #21267952 - 02/13/15 08:09 AM (8 years, 11 months ago)

:nodofunderstanding:


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