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Invisiblenatedawgnow
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SEMI-SOLID LIQUID CULTURE * 26
    #26689470 - 05/23/20 12:54 AM (3 years, 8 months ago)

:chief:SEMI-SOLID LIQUID CULTURE:chief:

Hello everybody and welcome to my thread!

So I've been working recently on a novel LC tek for funsies and for science!

I'm calling it semi-solid liquid culture (SSLC). So I noticed that LC tends to not like a rough shake,
and also tends to sink to the bottom of the jar/bag until colonized a bit for a shake, only at this point can it withstand
the damage from rough shaking. My solution to this problem? Enter SSLC :rockon:

The idea is to add a small amount of agar to your LC. I tested a bunch of ratios from .1-.5% and settled on .2% as the best ratio.
I used .5g agar for a 250mL liquid culture for this batch. This makes a very viscous soup. Anymore agar and it will solidify completely.
The amount of nutrients can be whatever you usually use for LCs. These were made with .2% nutes as well.

For this round I made 2 LCs, one traditional as a control, and one SSLC. So here are the 2 broths pre agar wedge so you can see how they look:

Regular LC


SSLC


Each LC was knocked up with a TOC clone.

Here are some pics a week or so into colonization. Unfortunately the SSLC is
kinda hard to see in the photos:

Regular LC


SSLC


Ok so I knocked up some jars with each LCs on the 14th. For each LC I made a a control jar where I just dumped it in
and let it sit, a shake jar that I shook vigorously, and a roll jar that I gently rolled back and forth to mix the
LC/grains. Each jar got around 10mL of LC, I free poured so impossible to be exact but that's what I was shooting for.

Here are the comparison shots between each LC on day 4. Roll and shake jars are from multiple angles. SSLC jars are on the left:

Control jars:


Roll jars:


Shake jars:


As you can see the SSLC jars are all almost fully colonized and the SSLC control jar shows a lot more growth than the
regular LC. My theory is that the myc has a stronger foothold because of the thicker medium and because of this it can
withstand more vigorous shaking and is less likely to sink to the bottom of the jars because it clings to grains.
My goal with it was to be able to shake it vigorously starting out to soak grains so that the jar colonizes in days instead of a week+.

As a side experiment, I dumped 50ml of SSLC into 1 jar to see how it will colonize. Here you can see how the SSLC
just sits on top of the grains until I shake it:


Here it is at day 4:


The jar is fully colonized, definitely a lot of moisture but no actual pools of water. This jar was shaken right after dumping in the 50ml.

Well that's about all he wrote! You may say "why bother when LC works just fine?"

My answer is because it's a hobby, it's fun, try some new shit every once in a while.

Let me know what you guys and gals think and if you have any comments please share!


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Invisibleeatyualive
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26689743 - 05/23/20 06:16 AM (3 years, 8 months ago)

:rockon:


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Re: SEMI-SOLID LIQUID CULTURE [Re: eatyualive] * 1
    #26690050 - 05/23/20 09:47 AM (3 years, 8 months ago)

Right on man I did exactly this with plates a while back during li experiments. 


It looks like this colonizes much faster than the plates did.

I’ll prob make one of these today myself.


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26690148 - 05/23/20 11:06 AM (3 years, 8 months ago)

Just used up the last of my LC so this is perfect timing! I'll give it a whirl


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: pureshrooming]
    #26690293 - 05/23/20 12:24 PM (3 years, 8 months ago)

Feel free to post pics and what not here when you do!


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26690793 - 05/23/20 04:41 PM (3 years, 8 months ago)

Just pulled 2 jars of LC with agar, 1 jar of standard LC, and 2 jars of water to make LI out of the PC. Gonna toss a wedge in all of them from the same clone plate and see who comes out on top! I'll update this when I have some results.


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: pureshrooming]
    #26690823 - 05/23/20 05:00 PM (3 years, 8 months ago)

Nice what ratios did you do for the sslc?

I found, and should have included im the tek, that with telephone agar it was
necessary to pre boil my broth to make sure all the agar dissolved before pcing.

Is yours nice and thick?


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26691321 - 05/23/20 09:55 PM (3 years, 8 months ago)

Yep nice and thicc! I do a pre-boil also so I got it nice and clear. It is definitely thicker than I expected but easily shaken.

I did 500g water, 1g agar, and 4g of lme then split that up between jars


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: pureshrooming]
    #26691394 - 05/23/20 10:37 PM (3 years, 8 months ago)

Make sure to give it a good swirl every once in a while to mix the colonized myc in.
Just don't get your filters wet with it, unless of course you're LC is filterless (way I do it)


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26692028 - 05/24/20 08:01 AM (3 years, 8 months ago)

I’m going to buy hay and hulls but when I come back I’m getting in on this.

I just can’t decide which type to do.

Probably albino cambo or APEU.


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InvisiblebodhisattaMDiscordReddit
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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26692144 - 05/24/20 08:53 AM (3 years, 8 months ago)

Nice write-up.

Agar in LC isn't very novel tho

https://www.shroomery.org/forums/showflat.php/Number/13781184/fpart/1/vc/1

And someone else recently made an adding agar to LC tek also saying it helped the LC stick to the grain giving faster recovery.
Ill see if I can find the thread it was the same exact concept.


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: bodhisatta] * 6
    #26692252 - 05/24/20 09:50 AM (3 years, 8 months ago)

If the only reference you can find is from 10 years ago I say it's relatively novel :shrug:

Nothing is entirely original anymore, that's just where we are in the evolution of the human mind


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26692277 - 05/24/20 10:00 AM (3 years, 8 months ago)

If you can find that other thread I'd be interested in reading it. I did a quick cursory
search from the past 5 years before posting the tek and pulled up nothing similar other than caps' sagari which is what gave me the idea


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26692291 - 05/24/20 10:07 AM (3 years, 8 months ago)

:takingnotes:


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Invisibleeatyualive
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Re: SEMI-SOLID LIQUID CULTURE [Re: staytrippy420]
    #26692316 - 05/24/20 10:16 AM (3 years, 8 months ago)

How fast of recovery improvement are you seeing? I think this would be great for bag culture. I feel LI and LC seem to seep through the cracks in bags a little more than jars. I’ve recently been getting 4 day colonization with LI using these new pour lids I bought. And this isn’t a variety that is generally fast. I also only did my initial shake after inoculating(#rubbedoneout)

Im also of the mind that the chunkier and thicker liquid inoculant is, the better it sticks to the grain and the faster it colonizes.

A lot of it has to do with the way you shake and roll jars. I usually fill my quarts really high for LI or liquids. I actually did a lower volume of inoculation than I normally do and it colonized in 4 days. 13 out of 19 jars did this. And the ones I didn’t shake quite as well are still at 75-90%. Ive since spawned those 13 quarts. This time I did 1 plate to 19 quarts where I normally do 1 plate to 10.




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Edited by eatyualive (05/24/20 10:52 AM)


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InvisibleLotKid
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Re: SEMI-SOLID LIQUID CULTURE [Re: bodhisatta] * 9
    #26692338 - 05/24/20 10:29 AM (3 years, 8 months ago)

Quote:

bodhisatta said:
Nice write-up.

Agar in LC isn't very novel tho

https://www.shroomery.org/forums/showflat.php/Number/13781184/fpart/1/vc/1

And someone else recently made an adding agar to LC tek also saying it helped the LC stick to the grain giving faster recovery.
Ill see if I can find the thread it was the same exact concept.



Looks to me like someone is unfamilair with the saying... "those who live in glass houses; probably shouldn't throw stones."


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Offlineverum subsequentis
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Re: SEMI-SOLID LIQUID CULTURE [Re: LotKid]
    #26692362 - 05/24/20 10:38 AM (3 years, 8 months ago)

Nice, Nate!


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: verum subsequentis]
    #26692476 - 05/24/20 11:34 AM (3 years, 8 months ago)

@eat- basically seeing full colonization between 4 and 5 days. Haven't done a bag
yet but that is next on the list.

@verum and lot- thanks guys :rockon:

"All that is old shall become new through rebirth."


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Edited by natedawgnow (05/24/20 02:05 PM)


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26692491 - 05/24/20 11:48 AM (3 years, 8 months ago)

Hah, I just inoculated a small jar of snot consistency agar like a few days ago.


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Edited by poisoned (05/24/20 11:49 AM)


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Invisibleeatyualive
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26692532 - 05/24/20 12:07 PM (3 years, 8 months ago)

Nice! I’d definitely be interested to see your results. I love liquids! :rockon:


Hmm snot agar you say....


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Re: SEMI-SOLID LIQUID CULTURE [Re: eatyualive]
    #26692587 - 05/24/20 12:40 PM (3 years, 8 months ago)

I like the sound of this...gonna give this one a go thanks nate!!


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: Zifozonke]
    #26692803 - 05/24/20 02:31 PM (3 years, 8 months ago)

Let us know how it goes!


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26701644 - 05/28/20 11:43 AM (3 years, 7 months ago)

I made one of these with natalensis the other day.

Did you notice if it took a little longer to colonize??

I’m feeling good about this one, the consistency is like chocolate syrup almost. Or watered down maple syrup.

I made a couple with different amounts of agar, the first one ended up solid lol.


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26701647 - 05/28/20 11:45 AM (3 years, 7 months ago)

No it colonized at the same rate as the other but the myc is more difficult
to see because of the opaqueness of the agar


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26701661 - 05/28/20 11:50 AM (3 years, 7 months ago)

Awesome.

I’ll probably pour Sunday or Monday and update.

This could definitely catch on, it’s the simplest and fastest method to get li which I’ve only done with ultra soft plates a couple times but it works great.


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26701672 - 05/28/20 11:53 AM (3 years, 7 months ago)

I like it purely for beimg able to shake the shit out of it and not have the inoculant
sink to the bottom of the vessel. To test these on agar, if you feel the need to,
just dip an inoculation loop into the sslc and streak it to a plate.

I haven't used a needle with these but it might be harder than just dipping a loop.


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26701679 - 05/28/20 11:56 AM (3 years, 7 months ago)

Yeah with lc I usually shake the jars a couple times the first day and once the second day to keep it from pooling.


I used syringes at first but lately I just pour it in.


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26707100 - 05/30/20 01:02 PM (3 years, 7 months ago)

4 days in from dropping wedges in two jars and biopsy method on another.

Super hard to see the progress - mostly to do with the jars I use - but it’s going!



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Re: SEMI-SOLID LIQUID CULTURE [Re: pureshrooming]
    #26707135 - 05/30/20 01:18 PM (3 years, 7 months ago)

Yeah I’ve been wondering if maybe some food color would make it easier to see how grown it is, I can’t see anything in mine.

Planning on cooking up some jars tomorrow and pouring Monday, mine will be six days at that point.


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26707139 - 05/30/20 01:19 PM (3 years, 7 months ago)

Doesn’t mycelium eat the colour after a while?


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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26707144 - 05/30/20 01:23 PM (3 years, 7 months ago)

Quote:

A.k.a said:
Yeah I’ve been wondering if maybe some food color would make it easier to see how grown it is, I can’t see anything in mine.

Planning on cooking up some jars tomorrow and pouring Monday, mine will be six days at that point.



Do you have a stir plate? 6 days aint tht long even cor regular lcs. Sslc needs
to be shaken or stirred relatively frequently to break the clumps of mycelium and agar up.


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Re: SEMI-SOLID LIQUID CULTURE [Re: staytrippy420]
    #26709119 - 05/31/20 01:14 PM (3 years, 7 months ago)

Quote:

staytrippy420 said:
Doesn’t mycelium eat the colour after a while?



I think that's the idea here. To see how much colour was eaten.


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Re: SEMI-SOLID LIQUID CULTURE [Re: poisoned]
    #26709123 - 05/31/20 01:16 PM (3 years, 7 months ago)

I don’t have a stir plate but I always shake them pretty violently to break the myc off the wedge.

I put in some time with a flashlight angling the jar and I can definitely see the myc floating in there.

There’s also bubbles which makes it tough.

I’ve got big expectations for this thing.


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Edited by A.k.a (05/31/20 01:17 PM)


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26709235 - 05/31/20 02:10 PM (3 years, 7 months ago)

I had the exact same thought on food coloring! Almost dropped some in when I made the jars but felt like I might get laughed off shroomery :grin:

I did a hearty shake earlier and put a flashlight to mine. There is some good progress but I'm going to wait about a week before I put it to grain. I will have 13 tubs of PE going as of today and need to pump the brakes a bit.


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Re: SEMI-SOLID LIQUID CULTURE [Re: pureshrooming]
    #26711453 - 06/01/20 12:54 PM (3 years, 7 months ago)

Alright I just knocked up six quarts of natalensis with this stuff. Hopefully it’s clean.

This variety is super fast and aggressive and apparently pretty contam resistant. I’m assuming there’ll be some fuzziness by morning.

This is the germ plate at 12 days from streaking. I had taken a few wedges 1.5-2 days earlier and it grew over so quick you can hardly tell.




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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26711475 - 06/01/20 01:01 PM (3 years, 7 months ago)

You'll start seeing recovery within 48 hrs if it's clean and full colonization
in 4-5 days if you shake right after pouring


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26711496 - 06/01/20 01:10 PM (3 years, 7 months ago)

I worked up a sweat shaking those jars. :box:

Lately I’ve noticed much quicker recovery with regular lc.  idk if it’s a coincidence and I’ve just had fast cultures but I switched my recipe and started using them within ten days also and I’ve been seeing 24 hour recovery.

Some of my other lcs that were older would take up to five days sometimes.


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26716959 - 06/03/20 02:29 PM (3 years, 7 months ago)

Two days after pouring


This stuffs sticky alright.


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26729279 - 06/08/20 04:07 PM (3 years, 7 months ago)

So i did a blenderless LI 250ml water with .2g agar. I decided not to put anything in the jar to help with the shaking because the blender ball annoyed me the last time but i think it hurt my cause. Things are still going well but a lil slower than what i'm seeing here. Still faster than when i did this without the agar though.

The first 4 jars were only shaken after the pour. The second 4 i shook the next day as well. That seems to have not worked out as well unless by some coincidence all of those jars got a little less than the others. The one least colonized on one side (with the big white wedge growth) was the last to be poured and didn't have much liquid but got a shitload of the agar puck. The center is probably pretty solid.

I've also noticed this PE6 myc seems to be slower in general than the other things i've got going on.

7th day:


A little more light on the 'once shaken' jars:


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Re: SEMI-SOLID LIQUID CULTURE [Re: Roger Clemency]
    #26729597 - 06/08/20 06:23 PM (3 years, 7 months ago)

I'm confused, were those done with blenderless LI or sslc?


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26729610 - 06/08/20 06:26 PM (3 years, 7 months ago)

So is this a soft agar like sagari, or actually a liquid?


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Re: SEMI-SOLID LIQUID CULTURE [Re: Caps McGee]
    #26729611 - 06/08/20 06:28 PM (3 years, 7 months ago)

It's a standard lc recipe with .2% agar added. It's a viscous syrup consistency
once cool from the PC. Inoc just like regular lc.


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26729627 - 06/08/20 06:33 PM (3 years, 7 months ago)

I just used the agar thickening idea while doing blenderless LI. I think a few seconds in a blender would make it work real well.


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26729631 - 06/08/20 06:34 PM (3 years, 7 months ago)

.2% of 500ml being 1g agar powder? Interesting... and what do you feel is advantageous?  I'm sure its listed on the op: quicker colonization or?

Edit: nevermind, reread, I gorcha


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Re: SEMI-SOLID LIQUID CULTURE [Re: Caps McGee]
    #26729642 - 06/08/20 06:39 PM (3 years, 7 months ago)

Its in the op, ya. Basically it clings to grains, doesnt sink, colonizes faster, ime. You can shake it hard due to stronger foothold on thicker media and pour a pretty significant amount without fucking up moisture content too much. In the op I dumped 50ml in one jar with zero sinking issues.


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26729655 - 06/08/20 06:44 PM (3 years, 7 months ago)

All same ideas I had with sagari...  I opted stiffer at 4.5g agar to 525ml, but effectively becomes a liquid upon the initial shake... I'd be interested in seeing where the balance point is precisely... really depends on whether you're looking for a pourable, or something you can take wedges from still... NICE WORK
:amusedapplause:


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Re: SEMI-SOLID LIQUID CULTURE [Re: Caps McGee]
    #26729700 - 06/08/20 06:56 PM (3 years, 7 months ago)

My jars I did with natalensis are doing ok on day 7. Not as fast as I hoped but I think it was due to grain prep.

I’ve made three of these so far just eyeballing different amounts of agar but I need to weigh it cuz it seems like a few extra grains of agar makes a noticeable difference here.

So far the best way I’ve found to gauge the mixture is to shake it and then backlight it and watch how fast the bubbles move through it to the surface.


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #26729794 - 06/08/20 07:30 PM (3 years, 7 months ago)

My jars are on day 7 and just about done. So far I like it! I always pour too much LC and have pooling on the bottom but this fixed that right up.


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Re: SEMI-SOLID LIQUID CULTURE [Re: pureshrooming]
    #26770515 - 06/22/20 02:14 AM (3 years, 7 months ago)

Hey Nate, what brand of agar do you use? I’m using telephone and it’s usually stiffer per gram than other brands.

Myc colonized on solid inoculant (g2g, a2g,li) tends to have a quicker leap off than normal liquid culture. I wonder how much the semi solid state of the media affects recovery time on grain.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yeetusdeetus]
    #26797669 - 07/01/20 07:38 PM (3 years, 6 months ago)

Just tried this out, fantastic results.. thicker was a bitter better for me.


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Re: SEMI-SOLID LIQUID CULTURE [Re: vertygo]
    #26801251 - 07/03/20 10:53 AM (3 years, 6 months ago)

:hellyeah:

@yeet- I use telephone brand also


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #26801278 - 07/03/20 11:12 AM (3 years, 6 months ago)

Forgot to take some pics but just spawned a few jars using this method and Im happy to say it worked wonderfully..thanks nate!!:thumbup::thumbup:


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Re: SEMI-SOLID LIQUID CULTURE [Re: Zifozonke] * 1
    #27010366 - 10/29/20 01:16 PM (3 years, 2 months ago)

Nice.
I like to do blender less LI using 1.5% agar in my plates but want to move to LC because it can be stored for a long time. But this looks more interesting than regular LC.
Do you know for how long SSLC can be stored?


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Re: SEMI-SOLID LIQUID CULTURE [Re: nosf3r4tu]
    #27010551 - 10/29/20 02:33 PM (3 years, 2 months ago)

Read this literally as I was making LC.  Had just measured out my water and sat down for a schmoke.  Spooky good timing for this to pop up. Trying now.  Thanks for the tip broski.


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Re: SEMI-SOLID LIQUID CULTURE [Re: alaskappalachian]
    #27010636 - 10/29/20 03:24 PM (3 years, 2 months ago)

I put a little agar in all my LC after trying this. It's a great way to have a colonized jar in 6 days if you need.


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Re: SEMI-SOLID LIQUID CULTURE [Re: pureshrooming]
    #27011710 - 10/30/20 09:17 AM (3 years, 2 months ago)

It can be stored as long as a regular lc. Havent pushed the limits but I imagine
you could get away with some extended storage times.


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27342577 - 06/10/21 06:59 AM (2 years, 7 months ago)

I realize this is an :oldthread:, but have a question.  Has anyone used vegetable glycerine instead of agar for this?  Just for funsies i might give it a try, just seeing if its been attempted.


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Re: SEMI-SOLID LIQUID CULTURE [Re: kingboomer]
    #27342603 - 06/10/21 07:38 AM (2 years, 7 months ago)

Quote:

kingboomer said:
I realize this is an :oldthread:, but have a question.  Has anyone used vegetable glycerine instead of agar for this?  Just for funsies i might give it a try, just seeing if its been attempted.


-Kingboomer



I used starch jelly from the mung bean which worked very well due to not really needing to worry about clarity.

I bought the stuff and had to find a use for it 😁



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Re: SEMI-SOLID LIQUID CULTURE [Re: DERRAYLD] * 1
    #27342782 - 06/10/21 09:29 AM (2 years, 7 months ago)

Never thought about using other thickening agents but I'm sure they work!

Glad people use this method. I havent done lc in a while but I may make up a few this week :rockon:


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27342805 - 06/10/21 09:40 AM (2 years, 7 months ago)

Crack showed us this link in the Discord on Monday or something and I got 20 pints of this ready to PC today.  Can't wait to use it.  Results look great, honestly.  Thanks for figuring this out, man.


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Re: SEMI-SOLID LIQUID CULTURE [Re: ShroomerInTheRye] * 1
    #27342948 - 06/10/21 11:18 AM (2 years, 7 months ago)

Haha just posted this but actually I kind of did this sslc by accident. Had only my agar premix on hand (agar
Peptone dextrose), made these little vials and noticed they were already jelling...so I dumped out half out of them and replaced it with water instead before pcing
Now I'm left with some sslc vials and the results are looking great
Imo! Better than my honey vials. Bout two days in.
At ten MLs it probably won't take long before jI can try using these. Overall I feel going slightly solid was a happy accident. Seems to keep the myc spread out but still moveable and it can still be sucked up with syringe.


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27437873 - 08/21/21 11:14 PM (2 years, 5 months ago)

Do you have any guess about a diagnosis for SSLC that isn't viscous when it pours? It could be:
  • Colonized too long
  • Agar "structure" got broken too much by shear, on a stir plate
  • Too much agar
  • Too little agar

Side note: it would be interesting to make SSLC from carbomer (acrylic thickener like in hand sanitizer or hair gel). It would be perfectly clear and shear would not break it up like agar can be broken up. Gelatin also supposedly reforms its bonds after being broken up, unlike agar.


Edited by gone-pear-shaped (08/21/21 11:53 PM)


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Re: SEMI-SOLID LIQUID CULTURE [Re: gone-pear-shaped]
    #27472610 - 09/18/21 06:16 AM (2 years, 4 months ago)

Prepared a media bottle with .1% LME and .2% agar. Will be inoculating this evening and will post results.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Lostkeys]
    #27475370 - 09/20/21 08:54 AM (2 years, 4 months ago)

Nice how did it go? See a lot of people adding agar to lcs lately :rockon:


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27475516 - 09/20/21 10:55 AM (2 years, 4 months ago)

Quote:

natedawgnow said:
Nice how did it go? See a lot of people adding agar to lcs lately :rockon:



It's colonizing


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27590479 - 12/22/21 09:50 AM (2 years, 1 month ago)

Hey LC fans

It's been over a year since this thread was bumped. How's it going with SSLC? Is this technique still producing good results?  Anyone have observations or innovations to add to the discussion?


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Re: SEMI-SOLID LIQUID CULTURE [Re: DharmaForKarma]
    #27590484 - 12/22/21 09:54 AM (2 years, 1 month ago)

Still works amazingly well.... nothing more to add.


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Re: SEMI-SOLID LIQUID CULTURE [Re: DERRAYLD]
    #27590494 - 12/22/21 10:07 AM (2 years, 1 month ago)

This thread was bumped 3 months ago! Give it a shot and feel free to post your results here


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27590519 - 12/22/21 10:29 AM (2 years, 1 month ago)

Quote:

natedawgnow said:
This thread was bumped 3 months ago! Give it a shot and feel free to post your results here




Jeez…2020, 2021. It feels the same.

Thanks!  I made a batch last night with the recipe from memory. 4g agar per liter, right?  🤢

Making a new batch right now, with the actual recipe.


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Re: SEMI-SOLID LIQUID CULTURE [Re: DharmaForKarma]
    #27590530 - 12/22/21 10:42 AM (2 years, 1 month ago)

Haha ya 4g per L is double the amount! Good luck on the new batch hope it works for ya!


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27706667 - 03/24/22 09:33 AM (1 year, 10 months ago)

I'm a bit late on this, but i fucking love it.. made 16 jars with 200ml white rabbit clone SSLC. Jars  got hard shake after inoculation. First sign of recovery were in 24h.. great stuff, i dont see myself going back to regular LC again after one try on this

Day 4


Shake on day 5

Day 7


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OfflineDendrocopos
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27724639 - 04/07/22 02:00 PM (1 year, 9 months ago)

Hey dawg. How do you mix these up? I usually put some bolts in my lc and kinda swirl it around. It's kinda hard to do this with sslc. Also, i didn't notice what kinda lids u using. I thought about just using unmoded plastic lids and agetating the lc by besting the shit out of it by banging in up and down.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Dendrocopos]
    #27724910 - 04/07/22 07:09 PM (1 year, 9 months ago)

They swirl just fine if your sslc isnt so thick that the whole thing
solidifies but I also use media bottles with unmodded cranked down lids so I shake em as well.


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27724918 - 04/07/22 07:16 PM (1 year, 9 months ago)

Damn i knew I should've gone unmoded, but i didn't wanna choke it. Polyfil with LC makes me nervous. The consistency is fucking fascinating though, haha. I will report back with results. Thanks for the Tek !


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Re: SEMI-SOLID LIQUID CULTURE [Re: Dendrocopos]
    #27725079 - 04/07/22 09:30 PM (1 year, 9 months ago)

I too have been playing with this fellas

I just used the leftover 30-50ml from pouring plates and topped it back up to 500ml, leaving me with around roughly 0.15% agar, 0.2% nutrition in my bottle I figure

As my agar included yeast, clarity of the SSLC was especially a disaster, so visual inspection for cleanliness is completely out with this method. I guess the agar also fucks with the clarity, depending on exactly which agar. But LC should be tested properly on plates anyway and this just makes you have to do that, instead of taking a shortcut going straight to grain with a broth that is visually clear of any turbidity

The myc really seems to like the colonizing in this more solid structure than in a more watery solution, and when it is spun it seems to recover in a very robust fashion quickly afterwards. With normal LC after vigorous spinning or shaking it seems the myc is almost harmed to some small degree, not so with SSLC I found

So the test on plates proved clean and I will use a little when the time is right and report back with results. I don't make much LC because I am a hobbyist level guy and aspirating LC into syringes from an open bottle in an SAB will test anyone's nerve. But when I do, it may very well be SSLC from here on in

Thanks again for the writeup Nate :cheers:


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Re: SEMI-SOLID LIQUID CULTURE [Re: SingularFusion]
    #27725278 - 04/08/22 06:04 AM (1 year, 9 months ago)

This is awesome! I never would have thought to just toss in a small amount of agar into my LC. PROPS!


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Re: SEMI-SOLID LIQUID CULTURE [Re: Danny_k] * 2
    #27725364 - 04/08/22 08:01 AM (1 year, 9 months ago)

Just stopping in to say, :thanx: I've been using strictly SSLC for 1 year now and I don't even remember what it's like to a2g or g2g. I think that I use a bit more SSLC in my grain than most people but with 7-10 day colonization, who cares?

I drop a piece of colonized agar to 100mL of SSLC and use 25g of colonized SSLC to inoculate 1 mycoquart (600mL) of grain. The grain is usually fully colonized in 7-9 days depending on the speed of the culture and recovers from a shake another 4 or 5 days later and then gets spawned.

SSLC is better than traditional LC IMO because you get even colonization through out the jar and don't have to shake the jars to distribute colonized pieces of grain.

I have friends who agree that .2% agar is too much because it makes their media too thick but that percentage will probably change with the different agar used. IIRC, they are using telephone. For what it's worth, I've been using Living Jin agar powder since day 1 and haven't had any issues.

If I don't shake the jars regularly, the myc starts to colonize the surface and will turn into a thick puck that's is a pain in the ass to break up. I free pour my media and when a chunk of agar gets to the lip of the jar, media drips and gets all over the place. I use to use magnetic stirrers but I got tired of losing my stir bars to the grain jars when they got low.




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Re: SEMI-SOLID LIQUID CULTURE [Re: Doctor Mario]
    #27726356 - 04/08/22 09:54 PM (1 year, 9 months ago)

:takingnotes:


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27726360 - 04/08/22 09:56 PM (1 year, 9 months ago)

Dang, everyone is unmoded. I really need to get my ADHD under control. I read about one sentence of the Tek and immediately went for it. I think I am gonna repour. Any kind of lc is a bitch with polyfil.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Dendrocopos]
    #27743641 - 04/20/22 01:12 PM (1 year, 9 months ago)



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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27743645 - 04/20/22 01:15 PM (1 year, 9 months ago)

Don't cut corners dude. Just use agar. Different kinds of thickeners than agar Are digested by mycelium. I am not sure what's in that one, but it would probably get less and less solid as the myc would consume it.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Dendrocopos]
    #27743921 - 04/20/22 04:28 PM (1 year, 9 months ago)

Nice write up! Trying this this week.


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Re: SEMI-SOLID LIQUID CULTURE [Re: GarryJarcia]
    #27744806 - 04/21/22 07:45 AM (1 year, 9 months ago)

I tried to use the agar but when I do it seems to just make a ring of solid LC at the bottom and the majority of it is liquid still. Any suggestions?


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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27744809 - 04/21/22 07:47 AM (1 year, 9 months ago)

Use unmoded lids. Put a marble inside. Shake the fuck out of it. Maybe make the water hot when adding the agar.


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Re: SEMI-SOLID LIQUID CULTURE [Re: LukeTheDuke187]
    #27744814 - 04/21/22 07:49 AM (1 year, 9 months ago)

Quote:

LukeTheDuke187 said:
I tried to use the agar but when I do it seems to just make a ring of solid LC at the bottom and the majority of it is liquid still. Any suggestions?




Probably using too much agar. Mine doesn't do that.


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OfflineLukeTheDuke187
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Re: SEMI-SOLID LIQUID CULTURE [Re: Doctor Mario]
    #27744818 - 04/21/22 07:53 AM (1 year, 9 months ago)

i used .4g for 250mL


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Re: SEMI-SOLID LIQUID CULTURE [Re: LukeTheDuke187]
    #27744841 - 04/21/22 08:10 AM (1 year, 9 months ago)

Boil your agar solution before pcing it to make sure all of the agar
is melted. Sometimes that happens if you just mix the agar and cold water then pc immediately.

All the agar sinks, melts in The pc cycle and then re solidifies as a puck at the bottom.


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OfflineLukeTheDuke187
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27744843 - 04/21/22 08:14 AM (1 year, 9 months ago)

Ah gotcha, thanks. What I did was use an electric tea kettle and put boiling water into a mason jar and added the agar. Do you think it didnt get melted enough?


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OfflineHongosamongus
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27745999 - 04/21/22 11:46 PM (1 year, 9 months ago)

Has anybody tried to use Semi-solid liquid culture for Paneaolus spp., or is this a no-go?


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Offlinekarri0n
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Re: SEMI-SOLID LIQUID CULTURE [Re: Hongosamongus]
    #27750037 - 04/24/22 10:00 PM (1 year, 9 months ago)

Quote:

Hongosamongus said:
Has anybody tried to use Semi-solid liquid culture for Paneaolus spp., or is this a no-go?




It's probably better. Pan myceliumn is wispier than cubensis and less robust after a shake, and this protects the myc.


--------------------

Panaeolus Bisporus


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OfflineCrackatoa
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Re: SEMI-SOLID LIQUID CULTURE [Re: karri0n] * 1
    #27750080 - 04/24/22 11:18 PM (1 year, 9 months ago)

Pan Cyan Rainbow Eucalyptus fruits


--------------------

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Offlinehazyhorse
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Re: SEMI-SOLID LIQUID CULTURE [Re: Crackatoa]
    #27750109 - 04/25/22 12:18 AM (1 year, 8 months ago)

this is badass nate!! thanks for the write up. definitely saving this thread to try in future grows


--------------------
you're not the first to set foot here, just another
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OfflineCatsLoveHouseMusic
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Re: SEMI-SOLID LIQUID CULTURE [Re: hazyhorse]
    #27789394 - 05/23/22 11:59 AM (1 year, 8 months ago)

So I was thinking about trying this as my first LC experience, do you guys suggest this tek over others or what do you think?

I’m doing an experiment. Half jars with quality SHIP/Syringe Filters and half unmodded in 500ml media jars. Should the unmodded ones be like 1/3-1/2 full? And when PCing the unmodded ones, you loosen the cap and use tin foil as usual, like with agar?


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Offlinekarri0n
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Re: SEMI-SOLID LIQUID CULTURE [Re: CatsLoveHouseMusic]
    #27789688 - 05/23/22 05:06 PM (1 year, 8 months ago)

Does SSLC ever get crystal clear the way a fully colonized broth gets?

I like that particular indicator in case of bacteria, and so far all of my SSLC mixes have stayed pretty turbid.


--------------------

Panaeolus Bisporus


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OfflineCrackatoa
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Re: SEMI-SOLID LIQUID CULTURE [Re: karri0n]
    #27789837 - 05/23/22 07:07 PM (1 year, 8 months ago)

I love this tek over any other LC recipes. I would suggest testing before using like any LC but the cloudiness makes it harder to read from the naked eye.
I only ever make 100ml-200ml of LC I knock up plenty with that and G2G from there if I want to. Unmodded just leave a little loose, cover with foil,PC and I tighten before even getting it out of the PC.


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Offlineholybrown101
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Re: SEMI-SOLID LIQUID CULTURE [Re: Crackatoa]
    #27789874 - 05/23/22 07:35 PM (1 year, 8 months ago)

no filter you say....
I'm knocking up 2 jars today but with jello, i have agar but wanted to try something different. Will post results. CANT WAIT


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OfflineCrackatoa
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Re: SEMI-SOLID LIQUID CULTURE [Re: holybrown101]
    #27789885 - 05/23/22 07:42 PM (1 year, 8 months ago)

Quote:

holybrown101 said:
no filter you say....
I'm knocking up 2 jars today but with jello, i have agar but wanted to try something different. Will post results. CANT WAIT



:llamastare::pondering:


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OfflineCatsLoveHouseMusic
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Re: SEMI-SOLID LIQUID CULTURE [Re: Crackatoa]
    #27790120 - 05/24/22 12:59 AM (1 year, 8 months ago)

Do you guys use .5G agar/250ml water? It says .2% nutes as well, but how much nutes do you use? I’ll be using LME.
Anyone add peptone and think it’s worth it?


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Offlinekarri0n
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Re: SEMI-SOLID LIQUID CULTURE [Re: CatsLoveHouseMusic]
    #27790765 - 05/24/22 02:56 PM (1 year, 8 months ago)

Quote:

CatsLoveHouseMusic said:
Do you guys use .5G agar/250ml water? It says .2% nutes as well, but how much nutes do you use? I’ll be using LME.
Anyone add peptone and think it’s worth it?




I followed the recipe exactly - 0.5g to 250ml the first time and ended up with a solid block.

I shook it and it's like the consistency of grape jelly now, a bunch of liquidy blobs and some chunks of solid agar in there. Not ideal, but it colonized ok.

I think depending on your brand of agar you will need to play with the ratios. 0.35 was better for me.


I also had one batch end up with a solid platter of agar on the bottom and liquid above. I know now to make sure to swirl it as it cools, and not let it cool to solid in the PC.

Everyone has a different ratio of nutrients they like in LC but from everything I've seen it looks like you can go realllllly low nutrients and still get a good amount of mycelium in a broth. 0.2% LME should be fine as would 0.1 or 0.5 or 0.05

I've never heard of peptone in an LC broth. Maybe in agar, but even then it doesn't seem necessary for any of the active species I've seen people cultivate or cultivated myself.


--------------------

Panaeolus Bisporus


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InvisibleMurphSmurf
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Re: SEMI-SOLID LIQUID CULTURE [Re: Crackatoa]
    #27790783 - 05/24/22 03:14 PM (1 year, 8 months ago)

Quote:

Crackatoa said:
Pan Cyan Rainbow Eucalyptus fruits





Bro ... hell yeah!


--------------------
Long-time lurker - love this site - love to learn - love agar - MMFam


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OfflineDendrocopos
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Re: SEMI-SOLID LIQUID CULTURE [Re: Crackatoa]
    #27790843 - 05/24/22 04:17 PM (1 year, 8 months ago)

Quote:

Crackatoa said:
Quote:

holybrown101 said:
no filter you say....
I'm knocking up 2 jars today but with jello, i have agar but wanted to try something different. Will post results. CANT WAIT



:llamastare::pondering:




:lol:


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OfflineCatsLoveHouseMusic
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Re: SEMI-SOLID LIQUID CULTURE [Re: karri0n]
    #27791298 - 05/24/22 09:54 PM (1 year, 8 months ago)

I appreciate the quality reply. All interesting real life experience that will definitely help. I’ve talked to a few people who absolutely swear by peptone, one said they feel like they grew incorrectly for years without it. IDK.

Do you use a magnetic stirrer?


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OfflineCrackatoa
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Re: SEMI-SOLID LIQUID CULTURE [Re: CatsLoveHouseMusic]
    #27791311 - 05/24/22 10:05 PM (1 year, 8 months ago)

I do, and you will with SSLC, IMO.


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Offlinekarri0n
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Re: SEMI-SOLID LIQUID CULTURE [Re: Crackatoa]
    #27793674 - 05/26/22 12:46 PM (1 year, 7 months ago)

I've never used an electric stirrer. I've only just started using SSLC, but I can say definitively, a very wide range of nutrients and nutrient levels work great with LC.




I've got psilocybe cubensis, Pleurotus ostreatus(oyster), Coprinus Comatus(shaggy mane), panaeolus Bisporus, and psilocybe natalensis on this shelf.

Nutrients in most of them are different, and different ratios from invisible to slight sediment to opaque(before colonization). The four that show cloudiness, starting three from the left, in this photo, are the SSLC.

Diluted Corn boil water, blackstrap molasses between 0.5g and 10g, WBS + Oat flour fully saturated water(the opaque one in the middle), half teaspoon of same flour water to 250 ml water, and LME at 0.1 to 0.2 % are all shown. Each colonized fine and most of them have been grown to fruits already.

I'm still trying to find a favorite nutrient and I am leaning toward flour water made from the grain you plan on using due to the recovery speed. I love the clarity of blackstrap molasses though. No sediment ever.


--------------------

Panaeolus Bisporus


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OfflineCatsLoveHouseMusic
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Re: SEMI-SOLID LIQUID CULTURE [Re: karri0n]
    #27794341 - 05/26/22 08:31 PM (1 year, 7 months ago)

I have LME and was thinking about using LME/Peptone/water in quarts and try some pints. Going to try some SSLC and regular.

I’m also going to try some jars with quality SHIPs/Syringe Filters, and some unmodded in 500ml media bottles.

Seems like people have so many different opinions, it’s tough to know where a good starting point is. I have to order the peptone still though, I also have PP5 plastic lids.


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Invisiblenix21
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Re: SEMI-SOLID LIQUID CULTURE [Re: CatsLoveHouseMusic]
    #27794498 - 05/26/22 11:10 PM (1 year, 7 months ago)

Did anyone tried store sslc in fridge for few months? Any inputs how it acts after extended period of time in cold storage?


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InvisibleSwabMarleyS
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Re: SEMI-SOLID LIQUID CULTURE [Re: nix21] * 1
    #27931607 - 09/03/22 01:57 AM (1 year, 4 months ago)

Bump!

Yes these store fine in the fridge just like a normal LC. Got one from 3-4 months ago that recovered on grain like it was freshly made!


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OfflinePadaone
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Re: SEMI-SOLID LIQUID CULTURE [Re: SwabMarley] * 1
    #27942556 - 09/09/22 07:43 PM (1 year, 4 months ago)

Great tek,

Gave it a go and seemed like I had clean growth. Nocc'd up some jars and its growth looks a little different to what I'm used to though looks similar to pics in OP, what do you guys think?



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OfflineCrackatoa
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Re: SEMI-SOLID LIQUID CULTURE [Re: Padaone]
    #27942574 - 09/09/22 07:58 PM (1 year, 4 months ago)

Quote:

Padaone said:
Great tek,

Gave it a go and seemed like I had clean growth. Nocc'd up some jars and its growth looks a little different to what I'm used to though looks similar to pics in OP, what do you guys think?





:asianofapproval:


--------------------

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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27942782 - 09/09/22 11:13 PM (1 year, 4 months ago)

Awesome Tek, I just wanted to post to keep it in my threads. Looks like I could use my tall pint jars for this SSLC with unmodified plastic lids I have. The tall pints don't work so well for me doing PF Tek. This looks like a great way to make use of them!


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Offlinejakemp
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Re: SEMI-SOLID LIQUID CULTURE [Re: Cob]
    #27956171 - 09/18/22 02:31 PM (1 year, 4 months ago)

This seems awesome! Will try this when i get to LC stage!


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OfflineScrewup
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #27976556 - 10/01/22 12:10 PM (1 year, 3 months ago)

Good contributions my guy 👍


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OfflineSymPlayTon
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Re: SEMI-SOLID LIQUID CULTURE [Re: Screwup]
    #27976604 - 10/01/22 12:35 PM (1 year, 3 months ago)

I made a batch yesterday.  Put a magnetic stir rod in it. 

It just sets up like a jello, but the stir rod was able to make it flow like a liquid still.  It is an interesting LC. 

I think an advantage it might have over Nirmal LC is the ability to fix air bubbles into suspension.  This creates a massive surface area, and gas pocket availability. 

I have yet to inoculate it still, but I will take pictures later and keep you updated.


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Re: SEMI-SOLID LIQUID CULTURE [Re: SymPlayTon]
    #27976619 - 10/01/22 12:42 PM (1 year, 3 months ago)

Let us know for sure! I am gonna try this within the next week or so. waiting for some agar plates to grow out.


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OfflineRusty2096
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Re: SEMI-SOLID LIQUID CULTURE [Re: SymPlayTon]
    #27977040 - 10/01/22 05:44 PM (1 year, 3 months ago)

Quote:

Cob said:
Awesome Tek, I just wanted to post to keep it in my threads. Looks like I could use my tall pint jars for this SSLC with unmodified plastic lids I have. The tall pints don't work so well for me doing PF Tek. This looks like a great way to make use of them!




Scroll all the way to the bottom. On the left. Click :wink:



Quote:

SymPlayTon said:
I made a batch yesterday.  Put a magnetic stir rod in it. 

It just sets up like a jello, but the stir rod was able to make it flow like a liquid still.  It is an interesting LC. 

I think an advantage it might have over Nirmal LC is the ability to fix air bubbles into suspension.  This creates a massive surface area, and gas pocket availability. 

I have yet to inoculate it still, but I will take pictures later and keep you updated.




Try with a little less agar maybe. I don't have a magnetic stirrer so I just dropped 3 pieces of broken glass in mine and I can easily hand swirl it.


--------------------
Currently looking for nothing. You guys who sent me stuff are straight up awesome!. :mushroom2:

We don't own things - things own us.

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OfflineSymPlayTon
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Re: SEMI-SOLID LIQUID CULTURE [Re: Rusty2096]
    #27977095 - 10/01/22 06:33 PM (1 year, 3 months ago)

Quote:

Rusty2096 said:
Quote:

Cob said:
Awesome Tek, I just wanted to post to keep it in my threads. Looks like I could use my tall pint jars for this SSLC with unmodified plastic lids I have. The tall pints don't work so well for me doing PF Tek. This looks like a great way to make use of them!




Scroll all the way to the bottom. On the left. Click :wink:



Quote:

SymPlayTon said:
I made a batch yesterday.  Put a magnetic stir rod in it. 

It just sets up like a jello, but the stir rod was able to make it flow like a liquid still.  It is an interesting LC. 

I think an advantage it might have over Nirmal LC is the ability to fix air bubbles into suspension.  This creates a massive surface area, and gas pocket availability. 

I have yet to inoculate it still, but I will take pictures later and keep you updated.




Try with a little less agar maybe. I don't have a magnetic stirrer so I just dropped 3 pieces of broken glass in mine and I can easily hand swirl it.





I think what happened with mine is evaporation. 

I started with .5g for 250ml.  It went down to about 210ml during PC. That is a big difference in the ratio at that volume.


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Offlinejakemp
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Re: SEMI-SOLID LIQUID CULTURE [Re: SymPlayTon]
    #27977396 - 10/02/22 12:25 AM (1 year, 3 months ago)

Make sure your cap to the container is lose.. I i lost about 100mls on my last agar run.. I loosened the cap a little more on my second and didnt lose any!
(could of got lucky i guess..lol)


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Offlinenakadash1
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #28269606 - 04/09/23 06:59 AM (9 months, 15 days ago)

Are these low nutrient concentrations (like 0.1%) something specific to this agar LC technique or is it just a matter of preference? I used to use 2-4% dextrose/LME, which I recently reduced to 1.5% which seemed like a substantial decrease already. Going to 0.1% is so far from what I'm currently using that I can't help but to wonder how the results could be similar. Is it wrong to assume that there must be some diminished amount of mycelium after full colonization between using a 0.1% nutrient solution vs a 1.5% solution? Even if its not 15 times less mycelium, how can a similar amount of mycelium colonize the jar with so much less available nutrients?

Also, the comment about adding 0.5% agar to the LC causing it to solidify (and others reporting solidifying LC with similarly low concentrations) is very confusing to me. I make my plates with 1.8% to 2% agar; and when I tried concentrations between 1.6-1.7%, the agar was very jello-like and was difficult to cut and especially hard to transfer because I wasn't able to poke it. Instead I basically had to try to use the scalpel as a spoon and pick the soft jelly-like wedge up. I just can't imagine how there could be agar that is so different from other types that it would cause LC to solidify when used at 0.5%. :confused:


--------------------
#1 Tek
With utmost care and precision,I work to achieve my fungal mission.
Sterilizing all my tools and space,to ensure not a single spore is out of place.
I carefully select the fungal strain and onto the agar, I make it rain.
With a steady hand and gentle touch, I inoculate the plate without a hitch.
Success is sweet when it's pure and true, and my agar plates are a testament too. A job well done, with no contamination, my fungi inoculation, a thing of  admiration.
JK NO
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OfflinePandaskis
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #28269618 - 04/09/23 07:15 AM (9 months, 15 days ago)

I wonder if you can sub the agar for gelatin, that way you can make it more solid initially and as the mycelium eats the gelatin it will become more watery. Wonder if its possible to spot contam in its more solid form before the mycelium turns it watery.

Just some crazy thoughts haha


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Invisiblemilkboy
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Re: SEMI-SOLID LIQUID CULTURE [Re: nakadash1]
    #28269629 - 04/09/23 07:27 AM (9 months, 15 days ago)

Quote:

nakadash1 said:
Are these low nutrient concentrations (like 0.1%) something specific to this agar LC technique or is it just a matter of preference? I used to use 2-4% dextrose/LME, which I recently reduced to 1.5% which seemed like a substantial decrease already. Going to 0.1% is so far from what I'm currently using that I can't help but to wonder how the results could be similar. Is it wrong to assume that there must be some diminished amount of mycelium after full colonization between using a 0.1% nutrient solution vs a 1.5% solution? Even if its not 15 times less mycelium, how can a similar amount of mycelium colonize the jar with so much less available nutrients?

Also, the comment about adding 0.5% agar to the LC causing it to solidify (and others reporting solidifying LC with similarly low concentrations) is very confusing to me. I make my plates with 1.8% to 2% agar; and when I tried concentrations between 1.6-1.7%, the agar was very jello-like and was difficult to cut and especially hard to transfer because I wasn't able to poke it. Instead I basically had to try to use the scalpel as a spoon and pick the soft jelly-like wedge up. I just can't imagine how there could be agar that is so different from other types that it would cause LC to solidify when used at 0.5%. :confused:




Nutes in LC is very much a preference thing. I find high nutritional LC becomes very.... thick over time, this with the addition of agar in a SSLC can cause some damn thick solutions if sitting for a while which is not my preference.

Feel free to try .5% agar but you wont like the results, .1% works great, .2% felt unnecessary thicker to me. I also believe agar strength can vary a little by brand.


--------------------
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Invisiblemilkboy
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Re: SEMI-SOLID LIQUID CULTURE [Re: Pandaskis]
    #28269632 - 04/09/23 07:27 AM (9 months, 15 days ago)

Quote:

Pandaskis said:
I wonder if you can sub the agar for gelatin, that way you can make it more solid initially and as the mycelium eats the gelatin it will become more watery. Wonder if its possible to spot contam in its more solid form before the mycelium turns it watery.

Just some crazy thoughts haha



That would defeat the purpose of using sslc


--------------------
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OfflinePandaskis
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Re: SEMI-SOLID LIQUID CULTURE [Re: milkboy]
    #28269634 - 04/09/23 07:28 AM (9 months, 15 days ago)

I mean perhaps you can use it before its fully liquified haha, and still semi solid as i said, crazy untested thoughts :P


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OfflineSmoothcat
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #28322845 - 05/17/23 06:53 AM (8 months, 8 days ago)

Gonna use my first LC today in a bag (always be baggin :thumbup:) the bag has been sat for the best part of a year in a tote. I just put it in the PC again last night as who knows how sterile it could be at this point. I think the grains have dried a little over time as well so hopefully the LC will add a little extra hydration

Anyway was looking at stuff for using LC and came across this thread so just wanted to get onto it and will probably be giving it a go after I pour out the other LC

Nice one Nate - I do like your teks :rockon:

Need to get my supporter status back though as cant post pics at the moment :sad:


--------------------
Back once again with the ill behaviour



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Invisiblenatedawgnow
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Re: SEMI-SOLID LIQUID CULTURE [Re: Smoothcat]
    #28322931 - 05/17/23 08:23 AM (8 months, 8 days ago)

Nice dude hope it goes well :rockon:


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Invisiblealtford78
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Re: SEMI-SOLID LIQUID CULTURE [Re: natedawgnow]
    #28339475 - 05/29/23 01:33 PM (7 months, 27 days ago)

How long do wait after inoculation to put it on the stir plate? 24-36hours?

I did two 200ml media bottles on Sunday, one with 0.1% agar and the other with 0.2%. The 0.1% is pretty much water like, the 0.2% is kinda syrupy but the stir plate manages to make an almost full vortex at 2000rpm.

0.2% LME ends up crystal clear, I might need to up that a little bit, I suspect my LME is on the weak end.

Awesome thread.:yougoodjob:


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28340518 - 05/30/23 11:11 AM (7 months, 26 days ago)

It's been almost 48h since inoculation, I put the stir plate for 10mins, came back and holy shit that's a lot of bubbles.

They are going to go away right? Right?:tryingnottodie:


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28340595 - 05/30/23 12:42 PM (7 months, 26 days ago)

Dunno - I dont use stir plates.  I swirl a bit each day (sometimes miss a day.)  I just didn't worry about it.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Bobgas]
    #28340610 - 05/30/23 12:54 PM (7 months, 26 days ago)

Quote:

Bobgas said:
Dunno - I dont use stir plates.  I swirl a bit each day (sometimes miss a day.)  I just didn't worry about it.



Same didn’t matter. I did mine in a media bottle and I’d shake that bitch like it was a grain jar


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Re: SEMI-SOLID LIQUID CULTURE [Re: Screwup]
    #28340668 - 05/30/23 01:40 PM (7 months, 26 days ago)

You just leave your lids cracked for ge?


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Re: SEMI-SOLID LIQUID CULTURE [Re: aPurpleCray0n] * 1
    #28340686 - 05/30/23 01:49 PM (7 months, 26 days ago)

Quote:

aPurpleCray0n said:
You just leave your lids cracked for ge?



Truthfully I’ve only done SSLC one time. I kept the lid cracked and had cling wrap around it just incase lmao. But yes.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Screwup] * 1
    #28340703 - 05/30/23 02:07 PM (7 months, 26 days ago)

I have mine tight like a fish's asshole. Can't be fucked to deal with loose lids right now, I'll probably crack it an a couple days. Or not, we'll see. I remember reading somewhere that one guy had it tight and it made no difference.

Man those bubbles aint going nowhere. I fucked up, shouldn't have left it on the stir plate and left. How the fuck am I going to see anything in this lol.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Screwup]
    #28340980 - 05/30/23 06:47 PM (7 months, 26 days ago)

Quote:

Screwup said:
Quote:

aPurpleCray0n said:
You just leave your lids cracked for ge?



Truthfully I’ve only done SSLC one time. I kept the lid cracked and had cling wrap around it just incase lmao. But yes.



Awesome, I’ve got a clone culture I’ve been working on I want to try this with


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Re: SEMI-SOLID LIQUID CULTURE [Re: aPurpleCray0n]
    #28341115 - 05/30/23 08:57 PM (7 months, 26 days ago)

If you heat the jar in a water bath, the mixture will loosen up and the bubbles can rise to the top more easily. How hot can the mycelium take without harm? 90F maybe? Precise warming would be easy to do with a sous vide cooker.

Or maybe we are witnessing the birth of... BLUBBLE TEK.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28341635 - 05/31/23 09:05 AM (7 months, 25 days ago)

I have my grain water sslc proofing out right now I like this tek.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28343962 - 06/02/23 02:24 AM (7 months, 23 days ago)

I actually put it back on the stir plate at a lower rpm for 40mins and all the bubbles migrated to the surface. I then shook it gently by hand and the bubbles went back under the surface. So there's an easy fix if you want to clear them. I can see the myc growing so I don't really mind them anymore.

Side question, I've seen people that have their LC on the stir plates like 24/7. Is there a point on doing this? Doesn't the myc need some time to rest?


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28343968 - 06/02/23 02:29 AM (7 months, 23 days ago)

it's just energy waste, I wouldn't do that at all. I work with lc/sslc without stirplate


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Re: SEMI-SOLID LIQUID CULTURE [Re: Shrimps]
    #28343969 - 06/02/23 02:31 AM (7 months, 23 days ago)

Quote:

Shrimps said:
it's just energy waste, I wouldn't do that at all. I work with lc/sslc without stirplate



Yeah just like…swirl it lol.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Screwup]
    #28343976 - 06/02/23 02:45 AM (7 months, 23 days ago)

Yeah that's what I was thinking as well, what's the point of having them spin all the time.

I don't remember where I saw it, maybe its was one of the youtube pros :yields:

Dude had like a dozen stirplates just spinning. :shrug:


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28344273 - 06/02/23 10:08 AM (7 months, 23 days ago)

In yeast anyway, stirring definitely makes the yeast grow faster. But yeast is not mycelium. Maybe it doesn't help at all. But even if the stir plate has no effect on growth rate, it does keep the mycelium from clumping up, which is nice.

And, when you are making jars of LC, it's nice to spin the ingredients with a stir plate instead of a spoon.

I will keep using my stir plate since I have it and I like it, but I would agree it is not critical equipment.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28344371 - 06/02/23 11:49 AM (7 months, 23 days ago)

I have a plate and I like using too, I just was wondering why I read/saw something about having them spin all the time (or something like 10h a day). Isn't like even 5mins a day enough? or even 1min


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78] * 1
    #28344394 - 06/02/23 12:08 PM (7 months, 23 days ago)

How much spinning you need depends on what the mechanism of the benefit is, I guess.

For example when you are growing yeast for beer, they need oxygen to replicate. Spinning the culture increases the amount of oxygen that is dissolved into the solution.

But yeast need that oxygen mostly for creating the compounds that make up their cell walls. Mycelium may not be the same.

Well, let's look that up... OK after some googling, it looks like all fungi synthesize the same oxygen-containing compound for their cell walls. It's called ergosterol. (Other sterols are surely in the mix too, ergosterol is just dominant.)

It seems reasonable that mycelium, which also synthesize ergosterol, would benefit from dissolved oxygen the same way that yeast do.

So how much do you have to spin the culture to keep the optimum level of oxygen dissolved? Beats me, but there is probably enough data out there to figure it out.

But if you only do it 5 minutes a day, you may be undershooting the oxygen level for optimum growth... Whereas if you do it continuously, you are more likely to supply enough. Maybe you'll provide more than enough, but that seems harmless.

There are a lot of other variables... Temperature, surface area, permeability of whatever filter the LC jar uses, myc species...

But from what I know, if I am gonna spin it, I am gonna spin it gently but constantly. If I have more than one LC jar cooking, I let them take turns, switching jars every 8-12 hours when I walk by.

YMMV.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28344400 - 06/02/23 12:19 PM (7 months, 23 days ago)

Interesting, I will look into this, thanks!

Just so there is no misunderstanding, you have your LC spinning on 12h on/12h off cycles?

When you say "gently", how do you quantify gently?


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28344422 - 06/02/23 12:46 PM (7 months, 23 days ago)

I stir constantly but since I only have one stir plate, if I am doing more than one jar they take turns.

"Gently" means the vortex at the top is no more than about 1/2" deep. I never stir to the point that bubbles are introduced but everything is moving around freely. This is about 20-25% power on my Amazon stir plate.

Honestly I think you could get the same oxygen dissolving benefits with minimal surface motion. Look at a big aquarium, where the ratio of surface area to volume is worse... They stay oxygenated with relatively little surface motion. But stronger stirring keeps the myc from clumping up, too.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28344438 - 06/02/23 12:59 PM (7 months, 23 days ago)

Cool, thanks. I've spun mine at max rpm a couple times and yeah a lot of bubbles but the myc seems to recover pretty well.:shrug:


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78] * 1
    #28344445 - 06/02/23 01:06 PM (7 months, 23 days ago)

If I have a culture sitting off the plate and it clumps up, I give it a good hard spin to break it up before I try drawing it into a syringe. I agree the bubbles don't hurt in normal stirring... I just don't want to look at them, and you might hear the stir bar rattle at higher speeds too.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28346938 - 06/04/23 10:57 AM (7 months, 21 days ago)

are there any visual cues to indicate that the LC is ready to go to grain? I mean, at what point are you satisfied with the myc growth in the bottle?

also, if you want to test it, do you aspirate with a syringe or free pour as little as possible on an agar plate? I'm using unmodified gl45 lids so I'm just thinking of pouring on a plate.


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28346951 - 06/04/23 11:11 AM (7 months, 21 days ago)

To test I extract with a syringe. I did that through the SHIP (and it was the only use of the SHIP post-sterilization). But, I have also taken off the lid to get a needle into the LC and that worked OK too.

I cannot imagine freely pouring the tiny amount needed.


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28346964 - 06/04/23 11:27 AM (7 months, 21 days ago)

cool, thanks. Do you cool the needle in the LC ?

Yeah I was also wondering if it's possible to pour that little into a petri. will probably make a mess.

what needle gauge are you using? 14-16g should be fine I think, I have some long blunt nose 14g that should do nicely.


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28346978 - 06/04/23 11:38 AM (7 months, 21 days ago)

At the time I did this I used a new sterile 16 ga needle, so I did not have to heat it. 16 ga was bigger than needed and I didn't have anything smaller at the time, but it worked OK. Today I would use a 21 ga instead just to minimize wear and tear on the SHIP... Those are the two sizes I currently stock.

(I never flame-sterilize a needle if the needle needs to go into a SHIP or solution, I will just put on a new one. They are dirt cheap and that way I don't have to even think about cooling the needle. The heat also makes them less sharp, which sucks. If I am dispensing solution from the syringe without needing the syringe to touch another object, like dripping LC onto a plate, then I will torch it and cool it by squirting out extra solution. YMMV.)


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28347158 - 06/04/23 02:12 PM (7 months, 21 days ago)

Ime by five days it’s worth using. Definitely not as thick as it will be in another five days but I usually get fast growth even when it doesn’t look super colonized.


What I’ve been doing is using a ship and filling a 20 or 30ml syringe, dripping some on a plate, then capping the syringe up and once the plate shows clean using the syringe. Free pour works well too though.


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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
    #28347181 - 06/04/23 02:35 PM (7 months, 21 days ago)

Quote:

A.k.a said:
What I’ve been doing is using a ship and filling a 20 or 30ml syringe, dripping some on a plate, then capping the syringe up and once the plate shows clean using the syringe.




Good thinkin'!


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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
    #28350358 - 06/07/23 02:46 AM (7 months, 18 days ago)

Since the agar makes the LC turbid by itself, what kind of visual indicators do you use for bacterial contamination? The old "put something with letters behind the bottle and if you can't read it, it's bacterial" doesn't really work here.


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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
    #28361420 - 06/15/23 07:52 PM (7 months, 10 days ago)

Made 3 of these last night - it was late and accidentally put 4g lme and 4g agar in each jar :foreheadslap:
Ended up making Super Solid Liquid Culture

:tryingnottodie:


Anyway I put all that in one jar and in the fridge and will try and dilute that at a later stage. Made 3 more with .4g lme and .4g agar

Got a couple of cubes and one pan

The wedges look like little jelly fish suspended in the soup - I like it

Be interested to see how this goes

:awehigh:


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