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Invisiblehamloaf
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The Learning Curve of LC * 12
    #28774930 - 05/15/24 08:44 AM (4 days, 21 hours ago)

There exists a considerable divergence of opinions regarding the efficacy of various types of liquid cultures, prompting discussions on their merits and drawbacks. Some contend that it's challenging to visually ascertain the cleanliness of a sugar-based LC.  However, akin to evaluating the cleanliness of a vendor spore syringe, the cleanliness of an LC can be reliably assessed through methods such as agar or petri dish testing.

It is worth noting that the PF Tek method has demonstrated success numerous times, often utilizing vendor spore syringes.  In the same way, tens of thousands of clean LCs have been cultivated from such syringes with success contingent upon factors such as the quality of the spore syringe, adherence to proper procedures, and the use of appropriate tools, filters, and containers (tens of thousands have not).

Despite the prevalence of novice practitioners opting for spore syringes from vendors to propagate LCs, the transition from a clean sugar-based LC to subsequent stages can be compromised by subpar inoculation procedures when introducing the LC to grain jars or bags.  Subpar techniques at this stage can lead to contamination of both the LC and the grain spawn it's introduced to.

Personally, my approach typically involves transitioning from spores to agar.  Through this methodical process, I maintain confidence in the cleanliness of my LCs, emphasizing the importance of a tailored approach and a willingness to learn from mistakes as part of the journey towards proficiency.



(Nice. thick LC)







It is advisable, though, to proceed with caution when dealing with liquid cultures inoculated with spores, as they may harbor contamination until proven otherwise through inoculation on test substrates and subsequent confirmation of cleanliness.

Common visual indicators of contamination in LCs include but are not limited to, a turbid or murky appearance, the presence of stringy or suspended growth, lack of translucency after incubation, absence of growth, or unusual odors.

The appearance of a dense white mycelial mat on the surface of the medium indicates an excessive level of colonization within the liquid culture.  If left unchecked, this mat can extend to fully cover the surface, forming a rigid layer of cells that can inhibit the oxygen exchange crucial for mycelial growth.  Consequently, this condition typically leads to the compromised health of the culture, often resulting in either its demise or an increased susceptibility to contamination.

Below is a link to a very simple and effective LC recipe.
https://www.shroomery.org/forums/showflat.php/Number/25184353#25184353


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Offlinetripdawg420
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Re: The Learning Curve of LC [Re: hamloaf] * 1
    #28774938 - 05/15/24 08:51 AM (4 days, 21 hours ago)

:rockon:


--------------------
HUSTLER
How U Survive This Life Everyday Resourcefully
epic GT mono tub
http://www.shroomery.org/forums/showflat.php/Number/17277772

wbs tek
http://www.shroomery.org/forums/showflat.php/Number/11525679
coir tek
http://www.shroomery.org/forums/showflat.php/Number/11917410
results :thumbup:

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Offlinefiddle_head
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Re: The Learning Curve of LC [Re: hamloaf] * 1
    #28775919 - 05/16/24 05:05 AM (4 days, 55 minutes ago)

Thank you for this hamloaf, i am driving but am about halfway through so far.


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Invisiblehamloaf
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Re: The Learning Curve of LC [Re: fiddle_head] * 1
    #28776249 - 05/16/24 10:13 AM (3 days, 19 hours ago)

I was hesitant to upvote your post due to the mention of reading while driving.  :lol:


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Offlinevicepope
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Re: The Learning Curve of LC [Re: hamloaf] * 1
    #28776341 - 05/16/24 11:47 AM (3 days, 18 hours ago)

Lc is the big curve, slurry is the big curve.

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Invisiblehamloaf
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Re: The Learning Curve of LC [Re: vicepope] * 3
    #28776754 - 05/16/24 06:30 PM (3 days, 11 hours ago)

Making mistakes is how you learn not to make mistakes.


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Offlinetripdawg420
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Re: The Learning Curve of LC [Re: hamloaf] * 1
    #28776879 - 05/16/24 08:14 PM (3 days, 9 hours ago)

Quote:

hamloaf said:
Making mistakes is how you learn not to make mistakes.



:rockon: practice dont sit around waiting on one grow every day do shit :headbang3: shit way to ez


--------------------
HUSTLER
How U Survive This Life Everyday Resourcefully
epic GT mono tub
http://www.shroomery.org/forums/showflat.php/Number/17277772

wbs tek
http://www.shroomery.org/forums/showflat.php/Number/11525679
coir tek
http://www.shroomery.org/forums/showflat.php/Number/11917410
results :thumbup:

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InvisibleBlazer420
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Re: The Learning Curve of LC [Re: tripdawg420] * 2
    #28777025 - 05/16/24 10:46 PM (3 days, 7 hours ago)

Could OP discuss their method on extracting LC from jars into syringes, as majority of the time extracting the LC from the jar can be the vector for contaminates if done incorrectly.


--------------------
~ I used to get high on life, until I realized life was cut with morons ~
* You need 2 wake up and smell the music! *
- We are all computer data in a materialistic world -
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Invisiblehamloaf
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Re: The Learning Curve of LC [Re: Blazer420] * 1
    #28777393 - 05/17/24 11:00 AM (2 days, 19 hours ago)

Doing LCs by hand, I allow the mycelium to colonize the whole media broth right up until the mycelium is about to reach the surface and only shake once at the time of drawing the LC into syringes.  When using recycled needles, I flame sterilize the needle before sticking it into the medium.  With brand new needles, I do not flame sterilize.  I am very mindful of not touching the media vessel, or the syringe to anything while making LC syringes.  I have a flowhood, so all sterile procedures concerning aspiring LC into syringes is done in front of the flowhood.


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Invisiblestubb
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Re: The Learning Curve of LC [Re: hamloaf] * 1
    #28777417 - 05/17/24 11:37 AM (2 days, 18 hours ago)

:offthehook:
Hell yeah. Very encouraging and eloquently put. :thumbup:

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Offlinefiddle_head
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Re: The Learning Curve of LC [Re: hamloaf] * 2
    #28778669 - 05/18/24 12:43 PM (1 day, 17 hours ago)

Excellent job hamloaf, thank you for this contribution. It is important to make newcomers aware of the risks inherent in attempting LC without prior experience in other media such as agar. I would like to add that visual proficiency and expertise in context clues can be everything when dealing with nutrient media such as liquid culture or semi-solid liquid culture. I opt for the latter, using a .1% recipe for both nutrient and agar amendments, as per milkboy. Semi-solid liquid culture with or without a .1% agar addition can be effective, but I find it draws into the syringes much easier and there is much less sediment. Although I do not reccomend SSLC to beginners, I do find it effective.
During my forays into this arena, I have often mistaken a bacterial ¨puck¨ for the thick mat of mycelium that can form on the surface of the liquid. When this happens, it is prudent to swirl or gently agitate the mixture, and monitor for a turbid, murky appearance as per hamloaf´s direction. I have included a photo of .1% SSLC for reference.


Additionally, here is a reference shot of an infected liquid culture, ganoderma.


Pleurotus Eryngii on SSLC



Additionally, a bacterial lc might also show signs of color change, possibly turning it slightly yellowish-green or another hue. There could be some sediment of floating clusters of bacterial colonies forming, creating uneven, textured or striated patterns in the media.
I thought this would be a prudent addition to the thread and would like to bump it to increase visibility. I feel there are too many newcomers trying their hand at LC before they attempt to learn agar.


--------------------
lagm 2024 || lagm 2023 || Performance - 2023 || Things That Are Good
=============
If you want to use cups instead of petris, read this sauce
🌿Looking for free spores? My inbox is always open.🐒

:swab:The Koru:petri:
XI 🅃 🄴 🄰 🄼    🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿 XI

Edited by fiddle_head (05/18/24 01:27 PM)

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Invisiblehamloaf
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Re: The Learning Curve of LC [Re: fiddle_head] * 1
    #28778965 - 05/18/24 05:27 PM (1 day, 12 hours ago)

Thank you fiddle_head, and thank you for your contribution to the thread.  :mushroom2:

If sediment in your LC is not really your thing; there is a fix.  It is a bit of an older technique, takes some time, but it checks out/works. 

Make your broth, jar it up and PC for 15 minutes at 15 psi, then allow it to cool.  Once it cools, pour the broth into another jar through a coffee filter. 

The coffee filter will catch all the sediment.  Place the strained broth into your jar of choice, PC again for 20-30 minutes at 15-17 psi, let cool, then inoculate.


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InvisibleBlazer420
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Re: The Learning Curve of LC [Re: hamloaf] * 1
    #28779340 - 05/18/24 10:27 PM (1 day, 7 hours ago)

@Hamloaf which jar tek do you use for the LC jars? Do you use injection ports on the jars? Or filter discs? Or simply open the jars to draw up the LC into the syringes?


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~ I used to get high on life, until I realized life was cut with morons ~
* You need 2 wake up and smell the music! *
- We are all computer data in a materialistic world -
| Sometimes you have to lose yourself, to find anything |

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OfflineTheLongComma
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Re: The Learning Curve of LC [Re: Blazer420] * 1
    #28779350 - 05/18/24 10:49 PM (1 day, 7 hours ago)

I've had success with LCs in the past, mainly due to the Josex poke. That shit made it stupid simple... but now I'm wondering if it was all beginner's luck.

I've recently made the foray into P.Cyans and SSLC at the same time. The culture I'm dealing with likes grain water... and seemingly grainwater only.

The combination of grain water and SSLC makes it very hard for me to tell if the culture is happy or not. The usual signs of clarity, color, reduced murky-ness have all gone out the window.

I am used to cultures settling towards the bottom and building upwards - but it seems that this culture is blooming near the surface and when given a good stir, seems to stay near the surface with hundreds of little myc dots.

Here's a shitty photo of an LC that I really have no idea about:


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:badadvice:

Edited by TheLongComma (05/18/24 10:50 PM)

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Invisiblehamloaf
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Re: The Learning Curve of LC [Re: Blazer420]
    #28780545 - 05/19/24 07:36 PM (10 hours, 24 minutes ago)

Quote:

Blazer420 said:
@Hamloaf which jar tek do you use for the LC jars? Do you use injection ports on the jars? Or filter discs? Or simply open the jars to draw up the LC into the syringes?



Half pint wide mouth mason jars, plastic lids with synthetic filter disks, no gasket, no injection ports, 10cc monoject leur locking syringes, 14 gauge piercing needles.  You are correct.  The lids of the jars are opened to draw LC into syringes working right up in the face of the flowhood. 

@TheLongComma, I think that LC is contaminated.  By the looks of it, and by what you are describing here and in the Pans thread, it is by definition “turbid”.  Which is characterized by the LC’s cloudy appearance and suspended particles.  Also, the culture being suspended is a visual sign of contamination.  The mycelium of a healthy LC will begin colonizing at the bottom and stay there while colonizing upward towards the surface of the liquid.  The liquid will also be transparent.



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