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mushhead
Potato Devourer



Registered: 08/22/14
Posts: 2,218
Loc: Dimension H-231
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Re: Some notes about LCs [Re: Blue Helix]
#27031644 - 11/10/20 01:24 PM (3 years, 3 months ago) |
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Pastywhyte said:
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p9hu7 said:
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Nurkurzda23 said: hi for how long do you autoclave? or for how long would you autoclave 4 x 250ml? 20-30min or longer?
15-25 is what youreaiming for. Liquids sterilize quickly at 15 psi, 15 mins is good enough. I do 20.
I’ve never been able to sterilize any liquid media or agar for 25 min or less and not end up with bacteria later.
Exactly this is why I PC for 30-1hr sometimes just to make sure my LC isn't going bacterial on me.
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Blue Helix
bold hand


Registered: 02/02/03
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Re: Some notes about LCs [Re: mushhead]
#27031709 - 11/10/20 02:06 PM (3 years, 3 months ago) |
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Quote:
mushhead said:
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Pastywhyte said:
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p9hu7 said:
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Nurkurzda23 said: hi for how long do you autoclave? or for how long would you autoclave 4 x 250ml? 20-30min or longer?
15-25 is what youreaiming for. Liquids sterilize quickly at 15 psi, 15 mins is good enough. I do 20.
I’ve never been able to sterilize any liquid media or agar for 25 min or less and not end up with bacteria later.
Exactly this is why I PC for 30-1hr sometimes just to make sure my LC isn't going bacterial on me.
I sterilize my LCs for 30 minutes at 15 PSI, but I also bring the LC to near boiling first by cooking it in the microwave for 4 to 5 minutes. I wonder if by putting the LC jars deep in the PC water like Asura says, you could get away with not doing the microwave step? At any rate, I'd suggest 30 minutes might be a little faster than ideal if the LC going in there is cold (room temperature) and is not deep in the PC water, especially if your LC is a full liter or something close. But if you put it in there hot (near boiling), I promise you that 30 minutes at a full 15 PSI is enough for 500 ml.
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coversall
إِنْ شَاءَ ٱللَهُ



Registered: 06/06/20
Posts: 2,749
Loc: संसार
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Re: Some notes about LCs [Re: Blue Helix]
#27031731 - 11/10/20 02:19 PM (3 years, 3 months ago) |
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Nice thanks for the feedback. Forgot to mention one batch was PCd for 45 mins and the other ~20 mins. The only visible difference is a slightly darker colour on the one that got PCd for longer. I've read about PCing for times between 20-60 mins. Is the canalization of sugars an issue?
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mushhead
Potato Devourer



Registered: 08/22/14
Posts: 2,218
Loc: Dimension H-231
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Re: Some notes about LCs [Re: Blue Helix]
#27033394 - 11/11/20 12:52 PM (3 years, 3 months ago) |
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Quote:
Blue Helix said: I sterilize my LCs for 30 minutes at 15 PSI, but I also bring the LC to near boiling first by cooking it in the microwave for 4 to 5 minutes. I wonder if by putting the LC jars deep in the PC water like Asura says, you could get away with not doing the microwave step? At any rate, I'd suggest 30 minutes might be a little faster than ideal if the LC going in there is cold (room temperature) and is not deep in the PC water, especially if your LC is a full liter or something close. But if you put it in there hot (near boiling), I promise you that 30 minutes at a full 15 PSI is enough for 500 ml.
Ah, so I usually heat my LC to dissolve the LME and its pretty hot, not boiling before it goes into the PC. I'll give this a shot. bring it to a quick boil at the same time as dissolving my LME eliminates the microwave. EDIT: Is there some kind of pro to microwaving other than to bring it to a boil? Does say using microwaves instead of induction heating better?
Edited by mushhead (11/11/20 01:00 PM)
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Nurkurzda23
Stranger

Registered: 07/13/20
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Re: Some notes about LCs [Re: mushhead]
#27042041 - 11/16/20 10:41 AM (3 years, 2 months ago) |
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Hi
Are there continuations ore is it just mycelium on the top?
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Celestialexplorer1



Registered: 03/25/20
Posts: 1,803
Loc: Floating in a tin can
Last seen: 2 months, 30 days
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I used those same bottles for a good while. The injection ports fit perfectly in the top.
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 To spend just one moment in eternity
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Geinstein
Shroomery addict



Registered: 01/25/18
Posts: 1,784
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Injection ports on those bottles truly . . .sucks
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Nothing breads nothing
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D3_Myc
Weeb Trash



Registered: 05/06/18
Posts: 4,399
Loc: Year Zero
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Re: Some notes about LCs [Re: Geinstein]
#27042177 - 11/16/20 12:19 PM (3 years, 2 months ago) |
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Are there any good pictorial threads on what to look for in a LC, what molds or bacteria look like in a LC? I got no idea what I’m looking for
Pics from sat I need to update them

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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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Re: Some notes about LCs [Re: D3_Myc] 1
#27042188 - 11/16/20 12:28 PM (3 years, 2 months ago) |
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Quote:
Nurkurzda23 said: Hi
Are there continuations ore is it just mycelium on the top?

Looks like myc to me. You'll know better if you inspect the myc that's started to crawl on the glass from the floating colony 
Quote:
D3monic said: Are there any good pictorial threads on what to look for in a LC, what molds or bacteria look like in a LC? I got no idea what I’m looking for
Pics from sat I need to update them


I have started a thread LC Training Camp for exactly this reason and I'll be updating it tonight as well with tests I have going on for antibacterial broths, I'd like you to post in that thread as well, I'm basically collecting documents (pics) and data so we can know more scientifically what we're up against (to be able to know what we're talking about instead of having to guess)
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BrownBear
Warrior-Traveler



Registered: 06/05/20
Posts: 1,539
Last seen: 8 months, 24 days
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Re: Some notes about LCs [Re: Mateja]
#27046008 - 11/18/20 05:46 PM (3 years, 2 months ago) |
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I haven't been satisfied with the quart jar lc's. Too much water and the bigger jar is a little difficult to work with when performing extractions. I recently bought 3 inch leur lock needles and switched to regular mouth pint jars. The first four jars I used 250 ml of water and the same amount of nutritional yeast (3 pinches) that I was using in the quart jars. Tonight I am sterilizing five 200 ml lc's made with four pinches of nutritional yeast. So I am increasing my nutrition and lowering the amount of water. Hoping to grow more culture with less water.
I have been dissapointed with the recovery of both oat jars as well as brf cakes. I decided to extract 3x60 ml syringes of water from the quart jar lc's. This leaves me with 8x10 ml of lc. So far I have seen far better recovery and growth. Here are pics of the lc, the lc in a syringe without removing water and the lc after removing most of the water.
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nmd_myco
Obsessed Noob


Registered: 09/05/20
Posts: 198
Loc: PNW
Last seen: 1 year, 4 months
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Re: Some notes about LCs [Re: BrownBear]
#27049239 - 11/20/20 04:24 PM (3 years, 2 months ago) |
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my first LC after 2 days. learning a lot in this thread.
foam is from the stir plate. blobs are from agar, which had a bit of yellow/green food dye.
will be going to uncolored plates on that last transfer before LC.
Edited by nmd_myco (11/20/20 04:26 PM)
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BrownBear
Warrior-Traveler



Registered: 06/05/20
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Re: Some notes about LCs [Re: Blue Helix]
#27050424 - 11/21/20 12:49 PM (3 years, 2 months ago) |
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Blue Helix said:
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Iambrownbear said:
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PitcherCrab said: What brand of LME is that Demonic? Both my agar and LCs have been ending up with sentiment for as long as I can remember and it drives me nuts. The mushrooms don't care, but I'd love to make everything a little clearer if possible. Or if you have other tricks to get them clearer I'd love to heard them. Your plates are beautiful.
I use .5 grams of nutritional yeast in my LC's. I have setiment but after a few days at low speed on the stir plate the mycelium colonizes it all and it ends up crystal clear. Thats when you know its ready.
Yeah, I just bought that too. I don't know if it works better or not yet (haven't tested it), but it does add a little more sediment. I agree with you, though: when the sediment is gone, the LC is done!
One thing I should also mention is that it's kind of good to have the agar plates you are using have most of the same ingredients in them (at least have them share the malt). That way the mycelium has the right enzyme profile right off the bat then you transfer it, so it'll have the jump on any bacteria in there.
I have successfully inoculated ny lc's with ny agar cultures. I should know in a few days whether it makes a difference or not. If it works, do you think that adding a pinch of ny to my oat jars would have a similar affect with recovery speed and colonization rate?
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A.k.a
Stranger



Registered: 10/27/19
Posts: 16,816
Loc: Gaming the system
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Re: Some notes about LCs [Re: BrownBear]
#27050548 - 11/21/20 02:30 PM (3 years, 2 months ago) |
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I’ve got a king oyster lc that’s doing some weird stuff.
It looks clean and tests clean on agar but it’s super slow.
The only things I can think of are senescence or some kind of bacteria that stunts the myc but doesn’t show up on plates.
I knocked up some wbs last week and it took a few days to recover but now it’s just kind of fuzzing up on a couple grains and not spreading.
Any ideas?
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LAGM2020     
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Om Namo Shivaya


Registered: 05/03/15
Posts: 749
Last seen: 15 days, 4 hours
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Quote:
verum subsequentis said:

truth
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Verum aren’t you the one with the crazy modded half gallon jar and spray nozzle thing??
Such a great contraption.
yep. But i rarely use it anymore. Maybe I'll bust her out and give her another shot. Free pouring is just so much less hassle.

I've been wondering about making a similar flask design as this, what syringe are you using here? Is it autoclaveable?
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Culliganman360
Stranger



Registered: 06/05/19
Posts: 854
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Can I make lc from prepped grain water. ? Just dilute it something like munch did with grain water for agar
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A.k.a
Stranger



Registered: 10/27/19
Posts: 16,816
Loc: Gaming the system
Last seen: 13 hours, 28 minutes
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Yep.
Agar and lc seem pretty similar in that anything nutritious seems to work fine. Plus with grain water there’s that theory that it’s better to use the same food through the whole process.
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LAGM2020     
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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Quote:
Culliganman360 said: Can I make lc from prepped grain water. ? Just dilute it something like munch did with grain water for agar
I'd say that the difference between how much you dillutd GW will be huge if you're using it for agar compared to a broth since so far from my estimations myc need approximately 40times higher nutrient density on agar compared to in a liquid medium. But yes you can definitely use GW, I use the Rye boil off after I finish prepping the rye and then I dillute that water up to 20 times until I end up with a very clear broth. And ask AK said it could help speed up the jump of to the grain post inoculation. Gl
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Culliganman360
Stranger



Registered: 06/05/19
Posts: 854
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Re: Some notes about LCs [Re: Mateja]
#27052783 - 11/22/20 06:45 PM (3 years, 2 months ago) |
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I’m doing 10 lc jars 5 of em are 400 ml water to 100 ml grain (oat) water Fucked up the other 5 jars so those are 600 ml water to 200 ml grain water
 Think that’s diluted enough?
After reading your post matea I should probably do more like 80 ml grain water to 400 ml water You think ?
Edited by Culliganman360 (11/22/20 06:49 PM)
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Mateja


Registered: 07/14/16
Posts: 7,948
Loc: Here
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This is 1 part GW mixed with 19 parts water

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Nobler Hino
a dojo and a forge?!


Registered: 08/29/15
Posts: 1,780
Loc: Deep Ellum
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Re: Some notes about LCs [Re: Mateja]
#27053340 - 11/23/20 04:28 AM (3 years, 2 months ago) |
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Grain water should be pretty rich with nutrients. I use to overkill a while back with gw and brf or malt.
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   "The sacred mushroom takes me by the hand and brings me to the world where everything is known. It is they, the sacred mushrooms, that speak in a way I can understand. I ask them and they answer me.” – Maria Sabina
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