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InvisibleJHOVA
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Re: Absolute maximum amount of LC safe to use [Re: mushpunx]
    #24925332 - 01/19/18 10:27 AM (6 years, 11 months ago)

From eats slurry in a hurry
Quote:

15. After you have inoculated all of your jars and bags. You simply shake the jars for 20 seconds each. You want to distribute that extra water you poured into your grain jars as much as possible so that you don't have any issues after this. At first you will get a little scared because your jars will look like there is pooling water at the bottom once they have sat on the shelf a few hours. Be patient, wait it out. The idea is to have fully colonized jars in 3 days without a shake and it works.




From muda LI thread
Quote:

once all you have finished the last inoculation. take your jars out of the SAB and give each of them a good shake.




From munch’s bli
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Quote:

shake receiving jars until the agar is visibly evenly distrubited throughout the jar




Ill have not tried not shaking. I had no idea there was faster recovery leaving it be for a while.


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InvisiblemushboyMDiscord
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Re: Absolute maximum amount of LC safe to use [Re: JHOVA]
    #24925349 - 01/19/18 10:39 AM (6 years, 11 months ago)

We using lc.

LI is just myc water.  Slurries are already colonized networks and bottles are different. Its an all in one.

But with grain jars plus lc..  Wait until you see it recover. Thats when i shake em.  They will be finished a few days later at most.

If not?  Something got dirty. And i get skeptical about spawning.

Lc shows no mercy

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InvisibleTheMadHatter420
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Re: Absolute maximum amount of LC safe to use [Re: mushboy]
    #24925392 - 01/19/18 10:58 AM (6 years, 11 months ago)

I will have to give the no shake a try on my next run of LC.


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Re: Absolute maximum amount of LC safe to use [Re: TheMadHatter420]
    #24925396 - 01/19/18 11:00 AM (6 years, 11 months ago)

Shit i mostly do it to examine my grains top layer.

I hate not knowing where the molds at.

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Re: Absolute maximum amount of LC safe to use [Re: mushboy]
    #24925398 - 01/19/18 11:01 AM (6 years, 11 months ago)

I fail to see how lc vs li would differ in when and why to shake. Lc is made of grain flour and water and li is made of grain flour agar and water. :shrug: maybe im missing something in their applications that they should be used differently or maybe old methods need an update.


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InvisiblemushboyMDiscord
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Re: Absolute maximum amount of LC safe to use [Re: JHOVA]
    #24925407 - 01/19/18 11:03 AM (6 years, 11 months ago)

:shrug:

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Re: Absolute maximum amount of LC safe to use [Re: mushboy]
    #24925433 - 01/19/18 11:13 AM (6 years, 11 months ago)

I think it has to do with the mycelium  actually growing on Solid matter and not just liquid.


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Re: Absolute maximum amount of LC safe to use [Re: JHOVA]
    #24925597 - 01/19/18 12:18 PM (6 years, 11 months ago)

Quote:

JHOVA said:
I fail to see how lc vs li would differ in when and why to shake. Lc is made of grain flour and water and li is made of grain flour agar and water. :shrug: maybe im missing something in their applications that they should be used differently or maybe old methods need an update.



LI is live mycelium that utilizes water as the transfer medium.  LC is the cultivation of a live mushroom mycelium culture (if it's clean) grown in an artificial, or organic medium containing nutes. 

Once either is running on grains no difference is made upon when it's shook.  An LI won't require a shake because it doesn't colonize a media.  An LI will only need to be shook in order for the mycelium to be loosened from it's original growing medium and suspended in water, then transferred onto it's new medium.


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Re: Absolute maximum amount of LC safe to use [Re: hamloaf]
    #24925601 - 01/19/18 12:20 PM (6 years, 11 months ago)

nice...:takingnotes:

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Re: Absolute maximum amount of LC safe to use [Re: mushboy]
    #24925662 - 01/19/18 12:53 PM (6 years, 11 months ago)

Are you sure it doesnt colonize a media? I have observed li syringes that have huge colonies inside them after say a week. It looks like its quite alive in there to me. Since they (lc and li) both contain the same ingredients and initial inoculant source at what point does li become lc?

Im going to test no shake LI next time side by side with shaking to see what differences i can observe. Its worth trying if you guys say it colonizes faster.

I have read you (hamloaf) saying to gently roll jars after inoculating them with LC instead of shaking vigorously. Do you do the same thing with LI?

I just wanna know whats best and fastest. Going by the 3 teks above hearing about no shake has me ...shook


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Re: Absolute maximum amount of LC safe to use [Re: JHOVA]
    #24925676 - 01/19/18 01:04 PM (6 years, 11 months ago)

Quote:

JHOVA said:
Im going to test no shake LI next time side by side with shaking to see what differences i can observe. Its worth trying if you guys say it colonizes faster.




Sweet, looking forward to seeing the results! I was just about to ask who want´s to split the burden with me of testing this theory out with LC and LI, shaking vs no shaking, but I don´t have free jars or the time to do both. So I´mma do this experiment with LC soon so we can finally see what this all means.


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Re: Absolute maximum amount of LC safe to use [Re: Mateja]
    #24925708 - 01/19/18 01:30 PM (6 years, 11 months ago)

I'm a total noob, being at the point of having some nearly colonized shoeboxes, but that is it... still I've been annoyed at the use of liquid culture and liquid inoculant as synonyms.

I've been using liquid inoculant lately, with whole plates or partials of agar mycelium, dropped into sterile water or grain water. These are used right away or stored in the fridge. LC, on the other hand, as I understand it, is actually germinated or transfered to and grown like an agar plate, except liquid.

I think I have run into problems with amounts of liquid in recent jars with LI, especially with LI which was made with less agar grown mycelium. It seems from observation that the less mycelium is present per amount of water, the less should be added to the grain jars. In some of my half pints from the last round, there is obviously too much water, even though I used the normal amount from my last round. The LI from these was made with far less mycelium from agar...even though they were made from transfers.

However, my first and second round of jars from great white monster culture were made from the germination plate. Half of an approximately 4 inch diameter agar plate was used, as opposed to half of a 60mm petri for the others (lots more mycelium). The LI was stored in the fridge for a couple of weeks between inoculations, yet the second batch is doing as well as the first. With these GWM inoculations, 100% colonization has been happening around 5-7 days.

My grain prep was basically what is described in this thread - rinse, long soak, 10 minute boil, steam dry. The only difference I have noted is the amount of mycelium added to the LI, since the same conditions and sterile practices were followed for all jars.

As a result, I have concluded that the amount of clean mycelium from agar placed in the LI has a direct bearing on the colonization time. I always shake the LI rapidly after dropping the agar, and I always shake the grain jars after pouring the LI.

Obviously, there are other factors which could come into play, but this is simply my observation from my recent one session inoculation of sixteen jars from three different varieties of cubensis LI.


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InvisibleTheMadHatter420
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Re: Absolute maximum amount of LC safe to use [Re: JHOVA]
    #24925712 - 01/19/18 01:33 PM (6 years, 11 months ago)

Quote:

JHOVA said:
Are you sure it doesnt colonize a media? I have observed li syringes that have huge colonies inside them after say a week. It looks like its quite alive in there to me. Since they (lc and li) both contain the same ingredients and initial inoculant source at what point does li become lc?

Im going to test no shake LI next time side by side with shaking to see what differences i can observe. Its worth trying if you guys say it colonizes faster.

I have read you (hamloaf) saying to gently roll jars after inoculating them with LC instead of shaking vigorously. Do you do the same thing with LI?

I just wanna know whats best and fastest. Going by the 3 teks above hearing about no shake has me ...shook





LI left to sit is similar to pastys EZ LC tek. The water leaches the nutrients out of the agar, in effect turning it into a weak LC over time. So then you can get clouds growing from the nutrients leached out.


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Re: Absolute maximum amount of LC safe to use [Re: Spivkurl]
    #24925721 - 01/19/18 01:37 PM (6 years, 11 months ago)

My li syringes are left in room temperature and grow like crazy. Ill take a pic later to show you.

Yea exactly mh420. If left in the fridge its probably going to stay uncolonized.

I love new experiments.


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Re: Absolute maximum amount of LC safe to use [Re: Spivkurl]
    #24925723 - 01/19/18 01:38 PM (6 years, 11 months ago)

Quote:

Spivkurl said:
As a result, I have concluded that the amount of clean mycelium from agar placed in the LI has a direct bearing on the colonization time.




LI doesnt colonize its just suspended. LC colonizes. wut are you referring too?

liquid inoculate - the wedge is chopped up and in liquid 'form'
liquid culture - a tiny wedge(or biopsy) is used to inoculate the rich broth, expanding the tiny piece into something that can inoculate many quarts.

you can do the same with LI, but youd need a new culture per X amount of LI jars made per Y amount of grain jars. or whatever.

**more LI is more cultures. more LC is just expanding 1 culture into endless broths. thats better..

Edited by mushboy (01/19/18 01:42 PM)

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InvisibleTheMadHatter420
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Re: Absolute maximum amount of LC safe to use [Re: mushboy]
    #24925727 - 01/19/18 01:39 PM (6 years, 11 months ago)

I think he is talking about the colonization time when put to the grains.


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Re: Absolute maximum amount of LC safe to use [Re: TheMadHatter420]
    #24925731 - 01/19/18 01:41 PM (6 years, 11 months ago)

:suicide:

oops:shrug:

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Re: Absolute maximum amount of LC safe to use [Re: TheMadHatter420]
    #24925739 - 01/19/18 01:44 PM (6 years, 11 months ago)

"I use a SAB

Let me make my question easier.

My LC has a nice giant cloud sitting at the bottom of my LC jar. (Typical yes)

What I typically do is stir it up pretty good to let the cloud move to the top of the liquid and float.

While it's floating I suck it up in a syringe and then use the syringe for my grain jars.

I don't see how the pouring method works effectively.

Do you shake your LC jar and then pour some stuff into fresh sterile grain jars?"


I just give the LC a quick swirl to mix it up and then pour some into the receivers, same as if I was doing a G2G transfer.

When I make LC, and I think this is the best way to make LC, I start by blending my agar wedge in some sterile water in an Eberbach container.. making an LI.
Then I pour it into my LME broth in a flask, and set it on a magnetic stir plate to spin for a few days.

It typically is ready to use in a few days, and there is no solid blob of myc that needs to be broken up or sucked up. The mYC will seperate and settle yea, but just a quick swirl and it's ready to pour or fill stringes


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Re: Absolute maximum amount of LC safe to use [Re: mushboy]
    #24925757 - 01/19/18 01:51 PM (6 years, 11 months ago)

Is it correct to assume that a liquid culture needs to go from a 'liquid' state into a 'solid' state before it can start colonizing solid materia?

This is why LI jumps off on everything faster than LC because LI is already in a state where it´s anatomically ideal for colonizing solid material. This anatomy is of course not ideal for colonizing liquids, and that´s why you get faster take off when inoculating LC with LC rather then with an agar culture.

Correct me where I´m wrong I´m just inviting a discussion!


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Re: Absolute maximum amount of LC safe to use [Re: mushpunx]
    #24925767 - 01/19/18 01:54 PM (6 years, 11 months ago)

Making LC out of blended agar seems intuitively faster than waiting for LC to colonize 2 weeks from a biopsy or small wedge.

8 days ago i added water to a jar using EZLC at a quarter size the mycelium in the jar is minimal. And thats after 7 days to get to quarter size! I was actually just thinking there has to be a faster method to get LC than 2-3 weeks. Thanks mushpunx fir the confirmatiin on my initial thoughts on LI.


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