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Offlinebeeker
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Registered: 08/30/05
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Seeking Pros and Cons of different LI techniques
    #26674389 - 05/16/20 08:31 AM (3 years, 8 months ago)

So I've got my agar technique down and some colonized jars and plates to play with. I'm wondering about what people prefer in 2020. I've seen Munch's and Muda's teks and they seem easy enough, but I like the idea of filling a large syringe with water, sterilizing it in a PC, and injecting it into a shaken up colonized jar of oats with a SHIP. Then just sucking up as much water as I can and going to work on inoculating some bags. It just seems with the myc already used to the grain it would be faster and I would be able to continue this process indefinitely with one jar of colonized grain. I saw this tek posted earlier, but can't locate it now. Is this a subpar method if I have countless clean agar plates? Time difference a non issue? I'm working with a flowhood if that changes things at all. Thanks


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Invisiblemushhead
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Registered: 08/22/14
Posts: 2,218
Loc: Dimension H-231
Re: Seeking Pros and Cons of different LI techniques [Re: beeker]
    #26674457 - 05/16/20 09:33 AM (3 years, 8 months ago)

I sterilize my syringes using the boiling water method it has never failed for me, I have reused 10cc plastic syringes this way until they basically don't plunge properly any longer
(https://www.shroomery.org/forums/showflat.php/Number/26011681)
You should start a LC from your grain water when preparing rye or other grains - this will introduce the myc to nutrients it'll be eating for the entire grow which imo helps with faster colonization. I use Light Malt Extract when I am not prepping grains
(https://www.shroomery.org/forums/showflat.php/Number/26022486/fpart/1/vc/1/nt/43)
I use a full 10cc syringe of LC for 1 bag (2 1qt jars) of spawn, I evenly spread the LC around the grain to cover as much space as possible with your myc.

With agar I start a MS culture and allow it to take over the plate.
I take cuts of myc growth that I discern to be either quickly gaining space or is just over all performing well - these cuts get a new plate obviously.
After I have narrowed down genetics on my plates I allow time for full colonization of the plate.
I add one fully colonized plate to one jar of spawn for quick colonization.

When you have fully colonized jars you can then begin Grain to Grain transfers which is by far the fastest method of colonization I have seen.
You have one master jar in which you spread colonized grain from that jar to other sterilized spawn jars.
All the above done with a Flow Hood or SAB.
Hope I helped!


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Offlinebeeker
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Re: Seeking Pros and Cons of different LI techniques [Re: mushhead]
    #26674471 - 05/16/20 09:44 AM (3 years, 8 months ago)

Thanks mushhead! I have plenty of grain master jars and have been expanding them effectively via G2G. I just feel I'm missing out on something and like the idea of jabbing a plunger in tons of bags in one go. The reality is I have little need for that much spawn. I guess I'm just trying to grow in this hobby and looking for a worthwhile challenge that expands what I'm able to do.


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InvisibleRoger Clemency
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Re: Seeking Pros and Cons of different LI techniques [Re: mushhead]
    #26674480 - 05/16/20 09:53 AM (3 years, 8 months ago)

The only LI I have tried is eatyualive’s blenderless LI. Sterilized 250ml water in a qt jar with a protein cup shaker ball inside while I was PCing grains, dropped in a whole colonized agar disk and shook.

I meant to prep my grains a lil dry but actually ended up on the wetter side so when I poured the LI in there was a few jars with real wet bottoms. Some I left upright, others I tipped to the side for 2 days and a coating of grains trapped the liquid there when I tipped back and everything else colonized fast. The wet bottoms held the water on those jars while everything else went well.

When I spawned tubs I was able to shake the good grain apart and keep it separate from the less colonized looking wet grains, but I saved all the wets and did a little tub and it’s colonizing fast too. We’ll see but so far it’s worked well


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Invisiblemushhead
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Registered: 08/22/14
Posts: 2,218
Loc: Dimension H-231
Re: Seeking Pros and Cons of different LI techniques [Re: beeker]
    #26674492 - 05/16/20 10:00 AM (3 years, 8 months ago)

Agar is a pretty worth while challenge and will help you grow immensely.
Liquid Culture is also a challenge because water is easy to sterilize but also extremely susceptible to contamination.
I have read that APE along with many other PE varieties are tough to grow.
You could start to learn about sclerotia https://www.shroomery.org/forums/showflat.php/Number/17910649
Grow some P. Mexicana and get some philosophers stones.


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Silence: Giving you room to listen / Stillness: Giving you room to feel / Spaciousness: Just giving you room


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Offlinebeeker
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Registered: 08/30/05
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Re: Seeking Pros and Cons of different LI techniques [Re: mushhead]
    #26682191 - 05/19/20 11:21 PM (3 years, 8 months ago)

I've actually got some APE jars going right now. Any idea on what's different about the growing conditions it prefers over other cubes? I know its a bit slower than the rest of my jars, but that's the extent of my knowledge.

Also, the sclerotia look like a fun project as well. Any preference on ps. mexicana A or tampanensis or ATL#7? Truffles are all new to me.


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Offlinemushpunx
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Registered: 04/20/14
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Re: Seeking Pros and Cons of different LI techniques [Re: beeker]
    #26682519 - 05/20/20 03:55 AM (3 years, 8 months ago)

I don't suggest making LI by injecting water into a spawn jar and pulling it back out. You will be pulling any bacteria (or possibly mold) along with it. Even if the spawn looks clean.
LI should only be made with a culture that has been transferred on agar until it's clean.


I use an Eberbach unit (that's what they are made for). They are autoclavable blenders for sterile tissue. You can make one using Mason jars and the blade assembly from an Oster blender, but if you look you can get real ones cheaply. They are super expensive new, the glass and metal ones I have in these pics are $300 + $500 respectively, but I was able to find them cheap on eBay (I paid $20 for the glass unit and just bought replacement lids. The blade assemblies and gaskets are inexpensive), they will last for life and are super nice. They fit any Waring blender base, I was lucky and found a lab blender with a seconds timer for consistency.. you don't want to damage the myc, 3 seconds usually is enough.

 

I just sterilize the Eberbach with water in it, drop a wedge in and blend. Then it's good to go.

I also use them to make liquid culture, by sterilizing LC broth (I use Erlenmyer flasks with threaded caps, I drill a hole and pop an SFD circle under, or use poly fill ) at the same time as my Eberbach.
I blend my wedge , then pour into the LC broth, and set on a magnetic stir plate until it's colonized, usually in about 3 days. This way it doesn't come out stringy or have blobs that need to be broken up. No need for special lids, spawn jars with GE filters work fine. I like the flasks, can be found for 5$ on eBay :shrug:


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Invisiblemushhead
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Registered: 08/22/14
Posts: 2,218
Loc: Dimension H-231
Re: Seeking Pros and Cons of different LI techniques [Re: mushpunx]
    #26682867 - 05/20/20 09:03 AM (3 years, 8 months ago)

WoW mushpunx I need to get me one of those!
I've been really wanting to get into LI and more in LC than I already have been.
If I can get my hands on one of these during the virus times I'll be set for LI and LC experiments.
Thanks for the inspiration!

EDIT: I found that the bottle attachment with blade is about $300 rn on ebay :3


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Meditation Principles

Silence: Giving you room to listen / Stillness: Giving you room to feel / Spaciousness: Just giving you room


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Offlinemushpunx
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Registered: 04/20/14
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Re: Seeking Pros and Cons of different LI techniques [Re: mushhead]
    #26682955 - 05/20/20 09:57 AM (3 years, 8 months ago)

Yeah it might take a little while but if you keep looking they do pop up for cheap on eBay pretty often - people don't know what they are or don't realize the resale value. Fungi Perfecti has lids and parts.

You can also make one yourself though, you can buy a cheap Oster brand blender or just the blade assembly for one - the assembly parts are metal and can be fitted to the ring of a mason jar. Might need a little silicone to stop any leaks but it's pretty easy to do :thumbup:


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Offlinebeeker
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Re: Seeking Pros and Cons of different LI techniques [Re: mushhead]
    #26683040 - 05/20/20 10:43 AM (3 years, 8 months ago)

Thanks for the suggestions, mushpunx. Seems like with everything else in this hobby, it's best not to rush into things but listen closely first to those who came before.


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