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JohnBravo
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G2G, LC, agar questions
#26630395 - 04/26/20 05:21 PM (3 years, 9 months ago) |
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Wanted a bit of advice on my specific situation.
Have a lot of grows under the belt; have been growing on and off for the past 10 years. Getting back into it in the past 8 months after about a 4 year hiatus.
I'm a pretty meticulous bastard when it comes to sterile procedure, so I've really only had one contam in the past several years and Im still not sure of the source. Anyway.
I'm a pretty busy entrepreneur with more money than time at the moment, so I'm outsourcing a lot the steps. I know a guy in the midwest with a farm, so he's selling/shipping me pre-hydrated, pre-PCed rye bags and malt-based agar plates for a minimal price. So I'm working with bags right now.
Previously, my workflow was spores --> LC --> grain bags --> 66qt monotubs with coir. Yields are inconsistent, however, and so I'm starting agar work. I purchased a 12x24 cultivation flowhood recently.
This is the first time I'll be working with agar (should have a long time ago, I know). From all the reading I've been doing, my plan so far is this.
I have several 66qts I just introduced to fruiting conditions a couple days ago, so I'm planning on taking several clone samples from fruits and working on agar, then isolating till they're contam free and/or show uniform growth.
This is where I'm a little fuzzy concerning my next move. Since the bags are coming to me pre-PCed and sealed, I was thinking of either doing an agar clone LC or G2G. From what I've been reading, G2G seems like the better option. If not, let me know. Again, I've been working with LCs exclusively for the time Ive been doing this, and probably have around 30-40 grows with one bag contammed and thats it. Mayhaps Ive been getting lucky.
So as far as a step-by-step, I'd cut open several bags (one bag at a time obviously) in front of the flowhood, put in an agar wedge, impulse seal them back up, let colonize.
After colonization, cut open other rye bag(s) and use about a handful of colonized grains from the master bags to inoculate the new bags. Is there a good way to close off the master bag in between the sealing and cutting of bags getting inoculated? Or is it find to just fold over? I know not to put my hand over it. I know it'll be in front of the flowhood, but it still makes me nervous.
My main question just revolves around the individual steps and minute details of the procedure.
If there's a better way you can think of to do this given my situation with the already sealed grain bags, let me know.
There's a lot of different tek pages kind of strewn around the website that I've been reading, but if theres a resource I missed and you can point me to it, that'd be much appreciated anyway.
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JohnBravo
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Re: G2G, LC, agar questions [Re: JohnBravo]
#26632446 - 04/27/20 03:49 PM (3 years, 9 months ago) |
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Bump. Any advice or pointing to a resource would be awesome, thanks .
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mushhead
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Re: G2G, LC, agar questions [Re: JohnBravo]
#26634608 - 04/28/20 03:02 PM (3 years, 9 months ago) |
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When I've used bags to transfer agar wedges onto rye I'd just cut it open after wiping it down with alcohol, drop the wedge in, and reseal it with micropore tape or really any tape as long as it seals it completely. This all of course done inside my SAB because I had no flow hood, however I'm sure it's the same difference mate. Also just for your convenience I'd use a master grain jar instead of a bag to keep your master sealed up properly.
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JohnBravo
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Re: G2G, LC, agar questions [Re: mushhead]
#26636935 - 04/29/20 02:04 PM (3 years, 9 months ago) |
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Thanks for the input. Anyone else for advice?
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mushhead
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Re: G2G, LC, agar questions [Re: JohnBravo]
#26637055 - 04/29/20 03:24 PM (3 years, 9 months ago) |
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https://www.shroomery.org/forums/showflat.php/Number/24363426 https://www.shroomery.org/forums/showflat.php/Number/15158218/fpart/1
I hate to reference redit here but... https://www.reddit.com/r/MushroomGrowers/comments/8t7aqu/techniquelooking_for_tek_on_using_spawn_bag_in/ or https://mycotopia.net/topic/104176-g2g-in-a-sab-with-a-bag/
However seems like the consensus is you don't have to seal your bags as long as you're using an SAB or Flow Hood, also seems like you may be able to use zipties to keep your bags closed/sealed, again as long as you're using an SAB/Flow Hood. I would say using spawn bags for G2G is doable, but at what measure?
LC would honestly be my way to play with spawn bags. I wish I could help you more.
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Edited by mushhead (04/29/20 03:33 PM)
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Caps McGee
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Re: G2G, LC, agar questions [Re: mushhead] 1
#26637070 - 04/29/20 03:32 PM (3 years, 9 months ago) |
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I use zip ties to close after inoculation: unload the bags in flow and theres no need to preseal in any fashion... this requires you allow the bags to cool either in the PC, or leave the LFH running for several hours... I've seen some gourmet growers whi never turn theirs off... I have an entire tek series linked in my signature explaining my process in detail: I'm currently exploring LC with oyster cultivation and can say 8ts a great deal finicky compared to A2G... I use a soft agar that basically liquifies with the initial shake, reserving the pros of both solid agar and a liquid innoculent... ime/o, liquids are unnecessary unless you're in a huge hurry for a whole lot... liquids are great for culture expansion, keep in mind this means ANY culture... you can escape bacteria and competitor molds on solid 2D agar: liquids spread them just as efficiently as the intended culture, and rarely result in overcoming infection of the nature... that said, it seems you've experience with spore straight to LC and wouldbt see much issue... gooe this is helpful
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JohnBravo
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Re: G2G, LC, agar questions [Re: Caps McGee]
#26637312 - 04/29/20 05:39 PM (3 years, 9 months ago) |
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Thanks a lot guys, you're all awesome
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Genel1941
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Re: G2G, LC, agar questions [Re: JohnBravo]
#26637360 - 04/29/20 06:06 PM (3 years, 9 months ago) |
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My first time with agar I tried to pour hot agar in a SAB that did not last long so I finished that on the cabinet put them in PC wrapped with foil plastic Petri 100x15 my first time just had 9 at about 12 psi for 50 minutes I let the pressure go down on its own completely opened up PC took off the foil and every-one was a small ball of melted plastic. I apparently did it all wrong? Can somebody tell me what I have done wrong?
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mushhead
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Re: G2G, LC, agar questions [Re: JohnBravo]
#26637362 - 04/29/20 06:09 PM (3 years, 9 months ago) |
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Quote:
JohnBravo said: Thanks a lot guys, you're all awesome 
Thanks Caps!
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mushhead
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Re: G2G, LC, agar questions [Re: Genel1941]
#26637364 - 04/29/20 06:12 PM (3 years, 9 months ago) |
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Quote:
Genel1941 said: My first time with agar I tried to pour hot agar in a SAB that did not last long so I finished that on the cabinet put them in PC wrapped with foil plastic Petri 100x15 my first time just had 9 at about 12 psi for 50 minutes I let the pressure go down on its own completely opened up PC took off the foil and every-one was a small ball of melted plastic. I apparently did it all wrong? Can somebody tell me what I have done wrong?
PC your agar solution using pyrex glassware or jar and allow it to cool to 120f at least, pour in an SAB using clean practices such as gloves, masks, and wash your hands. Pour each dish shallowly and carefully making a single stack of poured plates. Allow to cool in SAB to set so condensation evaporates, seal plates with tape/plastic wrap. I suggest you start your own thread mate. You may get help faster.
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Caps McGee
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Re: G2G, LC, agar questions [Re: Genel1941]
#26637508 - 04/29/20 07:32 PM (3 years, 9 months ago) |
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Quote:
Genel1941 said: My first time with agar I tried to pour hot agar in a SAB that did not last long so I finished that on the cabinet put them in PC wrapped with foil plastic Petri 100x15 my first time just had 9 at about 12 psi for 50 minutes I let the pressure go down on its own completely opened up PC took off the foil and every-one was a small ball of melted plastic. I apparently did it all wrong? Can somebody tell me what I have done wrong?
You pressure cooked petri dishes which are not autoclavable... only plastic containers with bottoms and lids both marked "pp5" will withstand the heat and pressure of the sterilization process... pouring petris can be done in SAB, I HIGHLY suggest only pulling stacks if 5 fron the sleeve at a time... it's a bit of a PITA, but I did it with a 66qt tub fir nearly a year before assembling the beast
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mushpunx
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Re: G2G, LC, agar questions *DELETED* [Re: Caps McGee]
#26638193 - 04/30/20 03:28 AM (3 years, 9 months ago) |
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Post deleted by mushpunx
Reason for deletion: Redundant
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mushpunx
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Re: G2G, LC, agar questions [Re: mushpunx] 1
#26638214 - 04/30/20 03:49 AM (3 years, 9 months ago) |
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How many quarts are these pre sealed bags you are getting and how many are you planning on doing?
How I proceed from there would depend on that information.
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migraineur
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Re: G2G, LC, agar questions [Re: mushpunx] 1
#26638245 - 04/30/20 04:29 AM (3 years, 9 months ago) |
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G2G is great and it saves you so much time. You can store grain in the fridge for months until you're ready to use it to do more G2G or spawn to bulk.
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LogicaL Chaos
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Re: G2G, LC, agar questions [Re: JohnBravo]
#26638297 - 04/30/20 05:24 AM (3 years, 9 months ago) |
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Personally i would use agar then make an LC with the colonized agar the use the LC to get the grains colonize. But agar-grain-to more grain works well too.
Prehaps use some metal clothes oins to seal the bags temporarily under your flowhood?
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mushpunx
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Quote:
LogicaL Chaos said: Personally i would use agar then make an LC with the colonized agar the use the LC to get the grains colonize. But agar-grain-to more grain works well too.
Prehaps use some metal clothes oins to seal the bags temporarily under your flowhood?
There really isn't any reason to close the master up during G2Gs, it's the receivers you want to close up asap.
I personally think G2G via bag to bag is a pain. Even after you wipe the bags with ISO the outside of the bags is still a contamination vector, and it's harder to control the mouth of the bag than with a jar, especially when working with two of them. Also he's working with a 12 x 24" hood (like the new mobile hood I built) which means he will have to tilt both the bags to keep them in the flow. I'd say that's the only thing that is more difficult with a shorter hood.
With bags I like to make quart grain master jars and either go 1qt into a 5qt bag or split each qt between two 4.5 qt bags to make a 5qt bag for each tub.
Or I just inoculate 5qt bags with LC or LI.
Agar and flow hoods have learning curves but if he can make a clean LC it might make more sense to inoculate the presealed bags that way.
If it were me I'd open the bags to pour LC in and seal them back up. Or use syringe
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LogicaL Chaos
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Re: G2G, LC, agar questions [Re: mushpunx]
#26638432 - 04/30/20 07:24 AM (3 years, 9 months ago) |
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Good points. Never thought of using a syringe for LC to inject into a bag. Im guessing the guage is quite big for that?
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Caps McGee
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Cold storage can be done with any innoculant: it's going to boil down to YOUR preferences, YOUR work flow and YOUR space... I'd suggest agar regardless, and as few steps between agar and bulk spawning as possible to achieve your needs... if you're only running a few tubs at a time, liquids are really not necessary... soft agar>bag>bulk... I opt to use master pints to limit loss with dirty cultures, but I'm not seeing enough issues to warrant the extra step/vector for contamination, so I'll be attempting plate straight to bag once this bag of milo is gone
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JohnBravo
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Re: G2G, LC, agar questions [Re: mushpunx]
#26638617 - 04/30/20 09:22 AM (3 years, 9 months ago) |
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So he does 2.5lb of rye per bag and Ive been doing 3 of those bags per coir brick in a 66qt monotub.
This next run I plan on doing alot - Probably 20 bags at once.
Sorry, should have included that info first.
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mushpunx
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Quote:
LogicaL Chaos said: Good points. Never thought of using a syringe for LC to inject into a bag. Im guessing the guage is quite big for that?
Some people wipe the bag with ISO and inject right thru it into the bag and cover the spot up with tape, mircopore or whatever.
I don't really like that idea though, I think it's better to pour it in the top or squirt it in with a syringe and seal up.
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