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Making Liquid Culture * 45
    #26022486 - 05/30/19 08:27 AM (4 years, 7 months ago)

:stirthepot:MAKING LIQUID CULTURE:stirthepot:



LC should ONLY BE MADE from clean agar plates. this is a point of logic not a 'can it be done' statement.
for the sake of making the 10 millionth 'why did i fail?' thread, just take my word and DONT MAKE LCs WITH SPORES.





WHAT IS LC?

LC would be the growing culture inside a nutrient rich liquid. the liquid is technically called a broth. this is also the case with agar.
so if you see the word BROTH, it simply means the liquid you have prepared.
AGAR and LC are the same thing(broths). only agar..has agar:stoned:

WHY LC?
because mushrooms:shrug:

HOW TO MAKE THE BROTH
measure out desired amounts of water. i like doing ~250ml water per broth. you can make gallon LCs if you want:shrug:
i do smaller amounts because they colonize quicker and 250ml of LC can last a long time. also cuts back on how much media you expose while inoculating and so on..

INOCULATION TEKS
JOSEX BIOPSY POKE - mandatory reading:oldman:
WEDGE METHOD(tek TBA)

LETS MAKE THE BROTH

weigh out 1gram of LME(light malt extract..powder!) i like doing ~1g per ~250ml water.

add the powder first THEN add COLD water. this prevents clumping.

do NOT use hot water to make broths:nono: the hot water can cook the grain powder giving you lumps:facepalm:

shake it up good..

sometimes i stash the prepared broth in the fridge for awhile(after grain cycles run) but you dont have too.

cover with foil, back off the cap and pc 15min @ 15psi.

liquids sterilize easily. 30min+ can caramelize the sugars/wastes time.
i remove from the PC the moment psi hits zero and tighten the cap:thumbup:

other points
unused broths have long shelf lifes. ive let them sit for weeks and they are ready for inoculation when you are.
i use unmodified lids. i screw the lid on tight after inoculation. the culture is suspended in liquid and they dont need much GE anyway.

NUTE TYPES:
you can use many nute sources to make broths. as with agar, the type of nute will determine clarity:thumbup:

LIGHT MALT EXTRACT


BRF unfiltered


GRAIN WATER


ive really only made three types. light malt, brf and grain water.

LC DISCLAIMER
some people say lc is advanced or contamination prone ect.. IME this is not correct. however, LC can and will exploit any cracks in your cultivation armour.
grains prepped too wet? cultures not entirely clean? SAB technique not down?? youll grow nothing but green:sad:


real truth
honey/sugary LCs are so 1995
:begone:


Edited by mushboy (05/30/19 08:59 AM)


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Re: Making Liquid Culture [Re: SFS96] * 5
    #26022636 - 05/30/19 09:50 AM (4 years, 7 months ago)

:rockon:

i like using media bottles for LC because its easy to pour into grain jars.

the mechanics of inoculating the broth and then using the lc to pour into grain jars is the same as pouring petris or doing g2g/a2g. easy peasy


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Re: Making Liquid Culture [Re: ShaperDreaming] * 3
    #26022643 - 05/30/19 09:54 AM (4 years, 7 months ago)

ooops.

post it anyway! maybe i forgot something or youll explain it in a way that will connect with more people:shrug:


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Re: Making Liquid Culture [Re: micelio] * 6
    #26022670 - 05/30/19 10:03 AM (4 years, 7 months ago)

i just pour them and eye ball it.

250ml of lc is enough for at least 25 jars. 50 if you got skills. i usually get about 30ish but im lazy like that.

i dont shake after inoculation either. just let them chill. i need to know if i contaminate from the liquid or from technique. impossible to tell if you shake shit up right away.

this means you did it correctly




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Re: Making Liquid Culture [Re: micelio] * 1
    #26022740 - 05/30/19 10:41 AM (4 years, 7 months ago)

the only times ive had the grains get too wet is if i prepped them kinda wet:blush:

OR, if i use the lc before its ready(colonized). like this one.


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Re: Making Liquid Culture [Re: Camera93] * 1
    #26022753 - 05/30/19 10:46 AM (4 years, 7 months ago)

i dunno. never done it before.

but i made this little cake by dumping some lc ontop but it took awhile to colonize.


id imagine the syringe injected lc would be better suited but aspirating into syringes is a different tek and requires modded lids.


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Re: Making Liquid Culture [Re: micelio] * 1
    #26023572 - 05/30/19 06:05 PM (4 years, 7 months ago)

Quote:

micelio said:
Mushboy..
Do you use leftover lc to expand out to new batches or do you start out with a wedge from a new plate..





no. always start with fresh wedges.

im bias towards honey lc because i tried it a few times and i always failed. plus every noob tries it and its as cliche as prom night acne.


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Re: Making Liquid Culture [Re: shroomify] * 1
    #26046107 - 06/11/19 02:57 PM (4 years, 7 months ago)

:awesomenod:In about a week


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Re: Making Liquid Culture [Re: verum subsequentis] * 1
    #26046360 - 06/11/19 05:10 PM (4 years, 7 months ago)

Quote:

ShaperDreaming said:
I just swirl it like it's a wine glass.




That's all I do. A good shake sometimes too :wink:


Quote:

verum subsequentis said:
You can absolutely use half, reload the SAB, and use the other half. You can even use half, let it sit for a couple weeks and use the other half.




:whathesaid:

Just like agar bottles.


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Re: Making Liquid Culture [Re: sandman420]
    #26051031 - 06/13/19 08:25 PM (4 years, 7 months ago)

mason jars are not in the tek I recommend media bottles over jars:shrug:

Either way unmodded never caused me issues but I've only done a handful of lc runs(with unmodded) so not much experience I guess.


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Re: Making Liquid Culture [Re: newtomyc] * 1
    #26055287 - 06/16/19 06:20 AM (4 years, 7 months ago)

Quote:

newtomyc said:
Mushboy, how often do you stir the broth after inoculation?




Maybe once a day or every other day:shrug:


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Re: Making Liquid Culture [Re: Pastywhyte]
    #26133000 - 08/12/19 09:54 PM (4 years, 5 months ago)

I like it too.

Pouring untested lc unto wbs is so ..rustic:shrug:


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Re: Making Liquid Culture [Re: primelines]
    #26133391 - 08/13/19 08:02 AM (4 years, 5 months ago)

Sure:shrug:

Use whatever you have that can be pced. I like media bottles.


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Re: Making Liquid Culture [Re: primelines]
    #26133423 - 08/13/19 08:29 AM (4 years, 5 months ago)




:cheers:
although i only make ~250ml broths. kinda wasting the other space. maybe i should get smaller bottles for lc and save the 500s for agar:rolleyes:


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Re: Making Liquid Culture [Re: verum subsequentis]
    #26133572 - 08/13/19 10:32 AM (4 years, 5 months ago)

Quote:

nix21 said:
for you to see full colonization on 250ml LC?




about a week or two. depends. the poke takes slightly longer because you are starting with tiny fragments opposed to a wedge.

Quote:

verum subsequentis said:

It is said sometimes that a colony floating means your lc is fubared but i have seen evidence that suggests otherwise.




a floater is what im looking for:awesomenod: colonies that settle are usually fucked ime

a healthly floater


some lazy turd


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Re: Making Liquid Culture [Re: verum subsequentis]
    #26133784 - 08/13/19 01:00 PM (4 years, 5 months ago)

Quote:

verum subsequentis said:
i thought he was talking about a colony that is literally growing on the surface of the liquid. From my observation, floating vs sinking can be caused by a few things. Namely, contams in the liquid/ culture or amount of oxygen in the liquid/ culture.






Ah I misread. Yah I agree with you. Colonies on the surface usually are bad.

Like this..


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Re: Making Liquid Culture [Re: verum subsequentis]
    #26133822 - 08/13/19 01:31 PM (4 years, 5 months ago)

Might be. It's an old pic but the lc was toast if I remember.


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Re: Making Liquid Culture [Re: Tweeq]
    #26133835 - 08/13/19 01:44 PM (4 years, 5 months ago)

:creepylurker:

..that is clear. Thanks man!


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Re: Making Liquid Culture [Re: LtLurker]
    #26133876 - 08/13/19 02:14 PM (4 years, 5 months ago)

Also use long ass loops.



I dont use stir bars because I dont need lc under a deadline.


Edited by mushboy (08/13/19 02:33 PM)


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Re: Making Liquid Culture [Re: the_sonic16] * 1
    #26134223 - 08/13/19 05:30 PM (4 years, 5 months ago)

I dont put filters on my lids. Dont want to mod the media bottles cap.

Leave the lid loose and covered with foil. Just like making agar. Same exact process.


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