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Maruta
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Grain LC or G2G (concerning sterility)? 1
#12410418 - 04/17/10 05:56 PM (14 years, 3 days ago) |
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Assuming a flow hood was out of the question, which method of inoculating WBS jars would be superior in terms of contamination?
After doing some research, this is what it looks like to me: 1) G2G is generally faster and simpler, whereas Grain LC involves some extra steps and materials 2) G2G is more prone to contaminants, especially without a flow hood 3) Both methods will result in more spawn than can be realistically used (by most people)
What I'm inferring: 1) Grain LC is less prone to contaminants, especially if you use simple self-healing lids. It appears that if done correctly, the jar contents are never really exposed to air (still or not). 2) If using a glovebox, G2G will be more likely to go bad compared to Grain LC.
So would Grain LC be the better choice given the circumstances? 1) Using simple still air glovebox with armholes cut out and Tyvek sleeves/rubber gloves. Liberal use of Lysol and rubbing alcohol, flaming, usual needle sterile technique. (question is, is G2G likely to become contaminated, and how bad compared to Grain LC) 2) Using SIMPLE self-healing lids (no premade, purpose built lids/filters) made from drilled lids, RTV silicone ports. I have read that it can't be completely air tight because of the vacuum/suction created (difficulty in drawing out LC too) so pressure relief is necessary. Is this true, and if so what kind of relief port is best (simple, effective)? I have seen simple polyfill ones to ones with another RTV port and a polyfill filled syringe. Maybe even simple Tyvek layer between lid with RTV injection port and a polyfill stuffed hole? Doc's WBS tek involves a single hole with polyfill (injection port AND air exchange) and a layer of Tyvek. Would this work for a Grain LC lid? Maybe not as contamination resistant as the RTV lid (maybe a hybrid lid would work)?
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As a side question, is there any significant disadvantages in terms of using BRF cakes to spawn to bulk (coir)as opposed to WBS/grains? It seems to me that BRF cakes are generally more reliable (less contaminant prone), and given some LC will do well in terms of speed. It seems to me that grain prep is more complicated in terms of ensuring proper water content (which has to be learned with experience), but it is simpler in that it involves only grain (whereas BRF requires vermiculite along with the BRF). So main question: is grain worth the effort in fussing with water content when spawning to bulk (coir)? Or will BRF cakes suffice?
Tentative plan: 1) Inoculate a dozen 1/2 pint cakes multispore using Spongiform's plastic BRF method (1/2 pint PC'able containers rather than traditional mason jars) 2) Optional: Inoculate 1 or 2 1/2 pint jars of grain multispore for Grain LC 3) After cake colonization, birth 4-6 cakes (double stack, small shotgun terrarium). 4) Harvest, and select samples (most robust fruit bodies maybe) for isolating work or cloning 5) Break up remaining cakes and use as spawn for coir (basically, run a small monotub along with cakes) 6) Create Grain LC from isolate or clone 7) Use Grain LC to inoculate more BRF jars or make more Grain LC 8) Cease direct birthing (except maybe to collect more isolates) and focus on using Grain LC->Cake->Coir monotub
-------------------- All threads, replies, posts, and material made by this account are fictitious and not to be taken literally.
Edited by Maruta (04/17/10 05:59 PM)
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Cloneufc
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Re: Grain LC or G2G (concerning sterility)? [Re: Maruta]
#12410470 - 04/17/10 06:08 PM (14 years, 3 days ago) |
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Both are good but I would go with G2G. With LC, if something goes wrong, you'll lose the whole batch. With G2G you might lose a few jars but not all of them.
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teesionbear
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Re: Grain LC or G2G (concerning sterility)? [Re: Maruta]
#12410496 - 04/17/10 06:12 PM (14 years, 3 days ago) |
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G2G is quicker if you leave room in the jar to shake it and mix it throughout.
Grain LC is easier to store long term and less contam prone.
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myceleus_rex
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Re: Grain LC or G2G (concerning sterility)? [Re: Maruta]
#12410502 - 04/17/10 06:12 PM (14 years, 3 days ago) |
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I'd say grain LC which is FAR more reliable than a standard sugar LC. It's still pretty fast for colonizing and you can easily make 500ml in one go. That's a lot of jars.
Grain jars are superior if you have a PC. Contamination is not likely with proper procedure and good lids. If you can make a grain LC, you can do grain jars.
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Maruta
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Re: Grain LC or G2G (concerning sterility)? [Re: myceleus_rex]
#12410557 - 04/17/10 06:24 PM (14 years, 3 days ago) |
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Ah I see, thanks that is very helpful.
So grain spawn is superior to BRF even if its to be used as spawn for coir? Wouldn't the fact that BRF is being used as spawn kind of obviate the 'ease of mass prep' advantage that grain jars have?
My understanding (correct please, if I am flawed): BRF Pro: -Fast and simple -Very low possibility of contam
BRF Con: -Pain to prepare more than a dozen jars -Birthing in cake form is tedious, given all the jar prep involved
By using BRF cakes as spawn, you are preparing less than a dozen cakes and the cons are negated. On the plus side, you have a very low chance of contamination in the jar itself.
When looking at Grain:
Grain Pros: -Easy to prepare many many large jars at once -Allows for G2G
Grain Cons: -More contam prone, especially without a flow hood (I plan on a glovebox) -More experience and finesse required in successful use (water content is important, heard of entire batches being useless because of messed up water content)
It seems to me that using BRF cakes as spawn and small amounts of Grain LC as inoculate plays to the strengths of BRF tek while mitigating weaknesses. Grain LC could be made in very small jars (even baby food sized). In doing so, they colonize fast and the myc can muscle out contams before they get a foothold. It is easier to aspirate out for LC, and it produces an adequate amount (only need enough several cakes, no need for large jars of it).
The only advantage I can see in using grains as spawn is the ability to G2G, and given that a glovebox is going to be used, Grain LC seems to make more sense. There would be minimal PC use (only for Grain LC jar prep) required. I'm really shooting for efficiency and contams would represent an unacceptable waste. G2G seems to be too risky with a simple still air glove box.
(please correct any factual information or reasoning, I have no real experience and I'm just speculating based on lots of lurking)
-------------------- All threads, replies, posts, and material made by this account are fictitious and not to be taken literally.
Edited by Maruta (04/17/10 06:28 PM)
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myceleus_rex
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Re: Grain LC or G2G (concerning sterility)? [Re: Maruta]
#12410620 - 04/17/10 06:41 PM (14 years, 3 days ago) |
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No offense, but you need to read more, GLC and grain jars both require a pc. With proper sterilization, grain jars are no more contam prone than BRF. They colonize faster, as you can shake them. You can produce far more spawn far quicker and they offer vastly more inoculation points when spawning to bulk, which reduces contamination risk.
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scatmanrav
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Re: Grain LC or G2G (concerning sterility)? [Re: myceleus_rex]
#12410624 - 04/17/10 06:42 PM (14 years, 3 days ago) |
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GLC doesnt require a PC if you use a PF cake as your myc source.
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myceleus_rex
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Re: Grain LC or G2G (concerning sterility)? [Re: scatmanrav]
#12410636 - 04/17/10 06:44 PM (14 years, 3 days ago) |
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How can you wash a descent myc slurry off of a solid PF cake?
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myceleus_rex
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Re: Grain LC or G2G (concerning sterility)? [Re: Maruta]
#12410738 - 04/17/10 07:10 PM (14 years, 3 days ago) |
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OP, you suggested in your plan: 2) Optional: Inoculate 1 or 2 1/2 pint jars of grain multispore for Grain LC
GLC does not work like that. You need ample room to wash the colonized, shook grain with sterile water. A pint jar with 100ml colonized grain is about right, like the jar on the right.
Study Agar's Grain LC tek
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Maruta
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Re: Grain LC or G2G (concerning sterility)? [Re: scatmanrav]
#12410764 - 04/17/10 07:15 PM (14 years, 3 days ago) |
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I would like to echo myceleus_rex's question. Is there a way to get a usable myc slurry off of a cake? User Spongiform has some posts about breaking up BRF cakes in the jar (plastic container) via shaking (he recommends striking the jar on suitable surfaces). I guess you could extract a slurry that way?
-------------------- All threads, replies, posts, and material made by this account are fictitious and not to be taken literally.
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Maruta
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Re: Grain LC or G2G (concerning sterility)? [Re: myceleus_rex]
#12410777 - 04/17/10 07:19 PM (14 years, 3 days ago) |
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No offense taken, I appreciate the input. What I meant by more contam prone is in terms of G2G. It seems to me that removing the lid and performing G2G without a flow hood (in a glovebox) is a relatively risky procedure. Using a Grain LC or other inoculate would obviate the need for G2G though.
So I guess the question is: Is the simple performance of grains over BRF as a spawn (colonization rate, ease of prep, etc.) worth the difficulty in dealing with water content (soak/simmer, etc.)? This is assuming there is no difference in contam rate (using Grain LC rather than G2G in a glovebox).
-------------------- All threads, replies, posts, and material made by this account are fictitious and not to be taken literally.
Edited by Maruta (04/17/10 07:20 PM)
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Maruta
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Re: Grain LC or G2G (concerning sterility)? [Re: myceleus_rex]
#12410794 - 04/17/10 07:24 PM (14 years, 3 days ago) |
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Quote:
myceleus_rex said: OP, you suggested in your plan: 2) Optional: Inoculate 1 or 2 1/2 pint jars of grain multispore for Grain LC
GLC does not work like that. You need ample room to wash the colonized, shook grain with sterile water. A pint jar with 100ml colonized grain is about right, like the jar on the right.
Study Agar's Grain LC tek
I was referencing some of the GLC teks on the board.
Fahtster's: http://www.shroomery.org/forums/showflat.php/Number/5781218#5781218
Other users like Doc have similar posts with similar takes on jar size.
-------------------- All threads, replies, posts, and material made by this account are fictitious and not to be taken literally.
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scatmanrav
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Re: Grain LC or G2G (concerning sterility)? [Re: myceleus_rex]
#12410806 - 04/17/10 07:26 PM (14 years, 3 days ago) |
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I Quote:
myceleus_rex said: How can you wash a descent myc slurry off of a solid PF cake?
It should come off in the water fairly easy. I admit I havnt tried it myself..but I have seen a few members who have and it worked for them. I injected a very full grain jar once and shook it around..it didnt break up..but I still got some good myc water.
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myceleus_rex
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Re: Grain LC or G2G (concerning sterility)? [Re: Maruta]
#12410821 - 04/17/10 07:30 PM (14 years, 3 days ago) |
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Quote:
Maruta said: I would like to echo myceleus_rex's question. Is there a way to get a usable myc slurry off of a cake? User Spongiform has some posts about breaking up BRF cakes in the jar (plastic container) via shaking (he recommends striking the jar on suitable surfaces). I guess you could extract a slurry that way?
I know what you're talking about, but with shook grain, it brakes down right to the individual kernels and you wash the myc right off the surface. With BRF, it will only break down so far, and if you introduce water, you'll basically make a mush that you probably won't be able to suck up because it will be full of spawn material.Quote:
Maruta said: No offense taken, I appreciate the input. What I meant by more contam prone is in terms of G2G. It seems to me that removing the lid and performing G2G without a flow hood (in a glovebox) is a relatively risky procedure. Using a Grain LC or other inoculate would obviate the need for G2G though.
That's why I said use GLC
Quote:
Maruta said:So I guess the question is: Is the simple performance of grains over BRF as a spawn (colonization rate, ease of prep, etc.) worth the difficulty in dealing with water content (soak/simmer, etc.)? This is assuming there is no difference in contam rate (using Grain LC rather than G2G in a glovebox).
Yes
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myceleus_rex
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Re: Grain LC or G2G (concerning sterility)? [Re: scatmanrav]
#12410844 - 04/17/10 07:36 PM (14 years, 3 days ago) |
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Quote:
scatmanrav said: I Quote:
myceleus_rex said: How can you wash a descent myc slurry off of a solid PF cake?
It should come off in the water fairly easy. I admit I havnt tried it myself..but I have seen a few members who have and it worked for them. I injected a very full grain jar once and shook it around..it didnt break up..but I still got some good myc water.
It might work, but if you do it by Agar's tek, you can easily get 500-600ml of clean, dense slurry out of 100ml of colonized spawn.
As Raoul Duke said, "Anything worth doing, is worth doing right"
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scatmanrav
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Re: Grain LC or G2G (concerning sterility)? [Re: myceleus_rex]
#12412192 - 04/18/10 12:43 AM (14 years, 3 days ago) |
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True...and I agree that it is so worth it to learn to do grain.
I just like to throw things out there. I used to be the one experimenting to prove possible what everyone said was impossible
I also do that fahkster thing..scoop a scoop into a small jar..then shake it..then you can keep the small jar and keep going back to it..but it requires the sterility of G2G (opening the lid, ect). Agars tek is the most surefire way to be sterile.
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Citric
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Re: Grain LC or G2G (concerning sterility)? [Re: scatmanrav]
#12412241 - 04/18/10 12:52 AM (14 years, 3 days ago) |
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Original tek was done using PF Jars. I used the idea of making a mini jar of grain(Poor mans way of Isolating[Drip one drop of spore solution in the middle, do not shake, and the dominant substrain should take over, giving you a form of an isolate]) and then loading it with sterile syringe water, shaking it all to hell, and getting not only water that touched the myc, but the actual myc that broke off.
Either way works, just however the grain way works better and easier.
If I were to do it with a PF jar, I would use the same idea. Do not fill the jar, and only fill it maybe 1/4 the way full. Use a tyvek filter, and you should be able to shake the hell out of it. Keep in mind you will lose a lot more water with a PF jar then grain.
Edit: Here's the link. http://www.shroomery.org/67/Poor-Mans-Mycelium-Syringe-Tek
-------------------- Self Healing lid tek ** Update 10.17.17 ** Mini casing pictures: Pins to harvest Cup O' Shrooms Magash: I noticed my contams were in the shape of fingers Hyphae: Yes "Loss of moisture from the substrate" is not a casing trigger. My final Grow!
Edited by Citric (04/18/10 12:52 AM)
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scatmanrav
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Re: Grain LC or G2G (concerning sterility)? [Re: Citric]
#12412286 - 04/18/10 01:00 AM (14 years, 3 days ago) |
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Yep, thats what I was talking about. That tek was around when I was a newbie.
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Citric
Registered: 03/19/04
Posts: 4,490
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Re: Grain LC or G2G (concerning sterility)? [Re: scatmanrav]
#12412321 - 04/18/10 01:07 AM (14 years, 3 days ago) |
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Quote:
scatmanrav said:That tek was around when I was a newbie.
...was? =)
-------------------- Self Healing lid tek ** Update 10.17.17 ** Mini casing pictures: Pins to harvest Cup O' Shrooms Magash: I noticed my contams were in the shape of fingers Hyphae: Yes "Loss of moisture from the substrate" is not a casing trigger. My final Grow!
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scatmanrav
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Re: Grain LC or G2G (concerning sterility)? [Re: Citric]
#12412396 - 04/18/10 01:24 AM (14 years, 3 days ago) |
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Quote:
Citric said:
Quote:
scatmanrav said:That tek was around when I was a newbie.
...was? =)
Hey you only got about a month and a half on me, sucka!
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