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OfflineKonnrade
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Start a liquid culture: +5,000 aspiration points
    #4836361 - 10/21/05 09:43 PM (18 years, 3 months ago)

If I have enough fruits soon to spare, I'll send a free spore print (B+ or PF) to the first person who explains the reference in the title.

I would like to get started learning about liquid cultures, and I have read the faq's pertaining to it, but personal opinions and anecdotes, as well as some miscellaneous info, would be much appreciated.

I plan on using a slant (slanted test tube) for my first try, since I've  got at least one of them to spare that's sufficiently sized. I intended to stuff the top with non-absorbent medical cotton.

I was going to try for a potato based medium, or honey/karo mixture. I have no agar at the moment but will try to get it for later attempts.

What would everyone recommend for me? Do I seem to have a decent plan? Any pointers/comments on using the potatoe based mediums? Should I mix in some honey/karo with it?

Thanks in advance for any feedback. Much :heart:


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Invisiblejarroddupont
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Re: Start a liquid culture: +5,000 aspiration points [Re: Konnrade]
    #4836380 - 10/21/05 09:48 PM (18 years, 3 months ago)

I can't exactly answer your subject question. But I can gtell you what I use for LC's. I mix 50/50 malt/dextrose (1 tblspoon) with a cup of water. This is a little (lot) more then the teks say, but IME the myc has taken off like crazy!

Thought maybe that would help you out.

Jarrod~


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P. Cubensis Growth Parameters
"All mushrooms are edible, but some only once."
-- Croatian Proverb


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Offlinezacharymanko
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Re: Start a liquid culture: +5,000 aspiration points [Re: Konnrade]
    #4836383 - 10/21/05 09:48 PM (18 years, 3 months ago)

Yea aspiration points as in sims 2. Any way experimentation on a variable should be well enuff, My experience is that the honey will make a slightly different color mixture but nutrients will be more.


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OfflineKonnrade
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Re: Start a liquid culture: +5,000 aspiration points [Re: zacharymanko]
    #4836485 - 10/21/05 10:28 PM (18 years, 3 months ago)

zach wins the print. Assuming I get one when I choose to harvest.

I was thinking the honey would be good, I knew it was a good source of nutrition for mycellium. Would honey water alone work, or should I mix it with potato starch leached into water?

Oh, and I don't precisely remember from the faq's, I can inject spores into the LC as opposed to a live peice of flesh, or can't I?


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Offlinesmarties
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Re: Start a liquid culture: +5,000 aspiration points [Re: Konnrade]
    #4836516 - 10/21/05 10:39 PM (18 years, 3 months ago)

im kinda new to LCs too. all i did was put a lil honey in alot of water in a jar and boiled it a few times in the microwave. let it cool all the way and inject like half to 1 cc of spore solution into the honey water and incubate. i used my LC to inoculate some quarts of wbs. they were fully colonized in 6 days. :laugh: so i guess its werkin. they are pretty simple once you get the hang of it.


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OfflineKonnrade
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Re: Start a liquid culture: +5,000 aspiration points [Re: smarties]
    #4836553 - 10/21/05 11:19 PM (18 years, 3 months ago)

awesome, that sounds pretty viable then!

I'm going to boil some potatoes, strain them, eat them (yummy potato!), and add some honey to the mixture. Then I'll chuck it in a PC and heat on low pressure for a while. I'll knock up a test tube with some B+ and instead of using spores on my next jars as I had planned I'll inoc them with some LC.


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Offlineantioch
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Re: Start a liquid culture: +5,000 aspiration points [Re: Konnrade]
    #4836893 - 10/22/05 01:32 AM (18 years, 3 months ago)

I've always used Roadkill's tek for LC. It can be found here:

http://www.shroomery.org/forums/showflat.php/Number/4438226

Good luck. :thumbup:

-antioch :mushroom2:


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OfflineKonnrade
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Re: Start a liquid culture: +5,000 aspiration points [Re: antioch]
    #4836962 - 10/22/05 02:05 AM (18 years, 3 months ago)

thanks for the link!

After filling my test tube 60% with the potato broth and honey mixture I had enough left to fill a 1/2 pint canning jar. So, I put it in there, put the lid on loosely, boiled it with the lid on top of a pot of water, and then tightened the lid. So, I have a nice sterile vaccuum canned substrate for future use sitting in my fridge. I may just punch two holes in the lid (one for needle and a bigger one with a filter for gas exchange) and knock it up later on.

For my test tube I put two layers of aluminum foil. One was bent in to form a bowl, then on top I put some cotton (all I had was normal cotton, no nonabsorbent yet) and put another layer of tinfoil on top. Sadly the syringe just took the cotton in with it in the cotton fell into the substrate. I'm betting on that tube contaminating, but I guess I'll see. I patched the top back up by stretching cotton across the hole and taping it to the sides held taught. I then stuffed a little extra under the cord of cotton. Nothing is getting in there except gas molecules.


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Offline_OttO_
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Re: Start a liquid culture: +5,000 aspiration points [Re: Konnrade]
    #4836971 - 10/22/05 02:10 AM (18 years, 3 months ago)

Quote:

Konnrade said:
awesome, that sounds pretty viable then!

I'm going to boil some potatoes, strain them, eat them (yummy potato!), and add some honey to the mixture. Then I'll chuck it in a PC and heat on low pressure for a while. I'll knock up a test tube with some B+ and instead of using spores on my next jars as I had planned I'll inoc them with some LC.




I really wouldnt bother with the potato water. Just use plain water and honey. You can start the LC with anything - spores, a peice of mushroom tissue or a chunk of mycelium.

When making LC, I simply boil a jar and its lid (with a little hole punched in it) in a pot of water for ten mins, Remove it from the water with tongs leaving some water in the jar - about 3/4 full. Add the honey, take the lid out of the boiling water with tongs and put it on the jar. Cover with foil, and then stick it back in the pot of boiling water with a tight fitting lid on for 40 mins to sterilise. If there is too much water left in the pot when you put the jar back in, tip some out.

To innoc, I simply crack the jar and scrape some spores in on top of the water with a sterile knife, put the lid back on, and shake vigorously to aerate the mixture, then incubate. When shaking, put your finger on the foil where the little hole is in the lid to stop any liquid getting out. I put a little dot on the foil with a marker so I know where the hole is in the future.  :thumbup: :mushroom2:


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OfflineKonnrade
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Re: Start a liquid culture: +5,000 aspiration points [Re: _OttO_]
    #4836987 - 10/22/05 02:16 AM (18 years, 3 months ago)

I know honey would be enough, but I figure it can't hurt to have starches in there too. Starches need no digestion to be used as far as I know. Plus, I wanted to eat the boiled potatoes anyhow.

I'm thinking I'll use roadkill's method, but with honey. I like the idea of putting a glass marble in there to stir and break up mycellium.

I really dont have much of a clean room, just a bathroom that I clean up and sterilize the area around the countertop of. So, I insist on inoc via syringe, it just makes contams less likely. Especially since bathrooms are prone to mold spores.

Random question:
what's a ballpark estimate for the time to colonize a slant of honey substrate?


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Offline_OttO_
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Re: Start a liquid culture: +5,000 aspiration points [Re: Konnrade]
    #4837053 - 10/22/05 02:59 AM (18 years, 3 months ago)

I do all my inoculation in the open air on a kitchen bench top, I just make sure it is nice and clean, spray it with lysol, wash my hands, close all windows - and I hold my breath when I do it. Zero contams with this method to date. Of course if you have syringes, this is safer, I only use prints because I think making a syringe is just another step where contams can get in, and I live in Australia so getting one from a vendor isnt an option - customs are very strict.

I estimate 1.5 weeks for your LC to be ready.  :thumbup:


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OfflineKonnrade
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Re: Start a liquid culture: +5,000 aspiration points [Re: _OttO_]
    #4837065 - 10/22/05 03:13 AM (18 years, 3 months ago)

Thanks for all the info! Very helpful. 5 mushrooms for you!

I look forward to seeing how fast a jar inoced from LC will take off.


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InvisibleFooMan
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Re: Start a liquid culture: +5,000 aspiration points [Re: _OttO_]
    #4837233 - 10/22/05 06:35 AM (18 years, 3 months ago)

Quote:

_OttO_ said:

When making LC, I simply boil a jar and its lid (with a little hole punched in it) in a pot of water for ten mins, Remove it from the water with tongs leaving some water in the jar - about 3/4 full. Add the honey, take the lid out of the boiling water with tongs and put it on the jar. Cover with foil, and then stick it back in the pot of boiling water with a tight fitting lid on for 40 mins to sterilise. If there is too much water left in the pot when you put the jar back in, tip some out.






Why do you boil twice? Seems like an unneccessary step to me.


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Offline_OttO_
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Re: Start a liquid culture: +5,000 aspiration points [Re: FooMan]
    #4837331 - 10/22/05 08:55 AM (18 years, 3 months ago)

I submerse and boil the jar and the lid for ten mins to pre-sterilise both of them, then I lift the jar out of the water retaining some of the boiling water in it to add the honey.

The water is still boiling hot and partly sterile, and the water on the stove is still boiling away.

I add the honey, chuck the lid on and it goes straight back into the pot thats still boiling. The water inside the jar doesnt have to get up to temp as it is already hot and the water is already part sterilised. Probably not 100% necessary, I just feel this is better when only steam sterilising.


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OfflineKonnrade
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Re: Start a liquid culture: +5,000 aspiration points [Re: _OttO_]
    #4838585 - 10/22/05 04:08 PM (18 years, 3 months ago)

I see the logic there, no reason against using an extra precautionary step, especially when it only means a few more minutes of time and a couple arm movements.


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