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OfflineNo-Pilot-555
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Super cloudy LC. What did I do wrong?
    #28008955 - 10/21/22 08:25 AM (2 years, 2 months ago)



I tried my hand at making my first Liquid Cultures last bit (haven't inoculated them yet.)
The Yellowish ones are 600ml of water,
20 grams of honey,
1 gram malt extract,
And .5 grams of yeast.
The white ones are 600ml of water,
18 grams Corn syrup,
1.5 grams malt extract,
.5 grams of yeast.
I didn't do any stirring just added magnetic stir into them all (you can see it at the bottom in some of them.) and put them in the PC at 15psi for 30 minutes.
They all came out super cloudy and just don't look right.
Someone please help. Thank you in advance.

Edit: Just tried this,
600ml water
9 grams honey/corn syrup
2 grams LME
.02 grams yeast.
Still super cloudy but going to keep two of them anyway.

Update: After my most recent try they were still a bit cloudy. Upon further investigation I've also found out I'm using way to much nutrients as well.

I kept two of the most recent recipe, one of both CS and H. They were still pretty cloudy but better. I'm going to give it a few days then test for contam.
My most recent recipe that I've tried was,
600ml water
3 grams CS/H
.5 LME
No yeast.
Everything I did the same. A lot more clear but still slightly cloudy. Going to give it a few days then inoculate them!


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Edited by No-Pilot-555 (10/22/22 03:04 PM)

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Invisiblemilkboy
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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555] * 2
    #28008962 - 10/21/22 08:29 AM (2 years, 2 months ago)

They are cloudy cause you used a shit ton of nutrients lol, I like .5% LME comes out pretty clear


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Offlinefather_fungi
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Re: Super cloudy LC. What did I do wrong? [Re: milkboy]
    #28009002 - 10/21/22 09:18 AM (2 years, 2 months ago)

There's nothing wrong with cloudy LC provided the cloudiness is caused by the ingredients - which I would say is what you have there.

In order of contribution to turbidity (fancy scientific name for what you have there - cloudiness) in my opinion:

1. The yeast
2. The LME

I can't tell with certainty if the quantities you used were per 600ml or per 1200ml split across 2 600 ml aliquots.  In the first case, 600ml, that's pretty heavy on the sugars, and second case, 20g/1200ml, is about right.

And finally, I always like to mention in LC discussions, any agar formula becomes a LC formula if you omit the agar.


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: milkboy]
    #28009009 - 10/21/22 09:25 AM (2 years, 2 months ago)

Do you think the yeast I added is over kill? I wanted to try it because I've had a lot of success with my plates using yeast. Typically my plates come out cloudy but that's usually no issue. As per mentioned I used .5 grams of yeast. Should I try something more like .1?


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: father_fungi]
    #28009010 - 10/21/22 09:26 AM (2 years, 2 months ago)

Thank you for the response. This actually helps a lot!
I wanted to try both honey and corn syrup adding malt extract and yeast to both. I'm using 600ml of water. Should I try 2% rather then 4?.


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555]
    #28009011 - 10/21/22 09:28 AM (2 years, 2 months ago)

What do y'all think about trying 10 grams honey/corn syrup, 2 grams malt extract, .2 yeast?


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InvisibleKnateKing89
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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555]
    #28009054 - 10/21/22 09:57 AM (2 years, 2 months ago)

Please don't bump your own thread in the first 24 hours. If you need to add information there is an option to edit posts. Your LCs are cloudy because of all the sugar. I use very little nutrients in my LCs like a gram or two of LME to 500ml water and it works great.

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Invisiblebakedbeings
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Re: Super cloudy LC. What did I do wrong? [Re: KnateKing89] * 1
    #28009059 - 10/21/22 10:04 AM (2 years, 2 months ago)

500mL water
quarter teaspoon BRF
best L ive ever C'd


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: KnateKing89]
    #28009079 - 10/21/22 10:34 AM (2 years, 2 months ago)

Sorry, I've never used a forum before. I don't understand how I bumped my own thread but I'm assuming by reposting to it? Either way, my apologies.
Okay thank you!


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: bakedbeings] * 1
    #28009083 - 10/21/22 10:36 AM (2 years, 2 months ago)

Quote:

bakedbeings said:
500mL water
quarter teaspoon BRF
best L ive ever C'd




This, this is great lmao. I'm going to have to try this.


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Invisiblemilkboy
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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555]
    #28009102 - 10/21/22 10:53 AM (2 years, 2 months ago)

Quote:

No-Pilot-555 said:
Do you think the yeast I added is over kill? I wanted to try it because I've had a lot of success with my plates using yeast. Typically my plates come out cloudy but that's usually no issue. As per mentioned I used .5 grams of yeast. Should I try something more like .1?



Personally I think its overkill, can you use it ? Yes, some people do! Do you need yeast? No, totally up to you. Also pretty much any nutrient you can use for agar you can use for LC.

I like SSLC but thats not as clear, so just depends on your intentions really


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: milkboy]
    #28009112 - 10/21/22 11:04 AM (2 years, 2 months ago)

Thank you for the information! I just wanted to try yeast. If it didn't work out, I won't be using it in my LC'a anymore.

The reason I'm so worried about it being cloudy is because I dont know how hard it's going to be to spot contam in my LC's.
This is my first go around with Liquid Cultures so I'm not sure how it's going to go really. I figured it being clear would just make it easier. But it doesn't necessarily have to be crystal clear. Just clear enough for me to spot contam.
I'll still be doing quality check regardless.

I've also never tried SSLC. I may have to give that a go instead.


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Edited by No-Pilot-555 (10/21/22 11:04 AM)

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Invisiblebakedbeings
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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555] * 1
    #28009135 - 10/21/22 11:29 AM (2 years, 2 months ago)

regarding forum rules, there are sticky threads for new people to become acquainted

regarding LC, i recommend to start simple. its a beginner tendency to want to mix as many ingredients as possible thinking it will make things better. we already know grain is a complete food for mushrooms so why not use it for LC, agar

its ok for LC to be cloudy with nutrients because as the myc grows it will clear the liquid up. my grain LCs start very cloudy and in the end they look like this



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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: bakedbeings]
    #28009198 - 10/21/22 12:04 PM (2 years, 2 months ago)

Wow, all of that really helps. Thank you so very much! I'll be inoculating my jars here in a few days (once I do quality checks on them and all.)
Again thank you!


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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555] * 1
    #28009270 - 10/21/22 12:54 PM (2 years, 2 months ago)

Bakedbeings said it best, keep it simple. Also clear LC is nice but visual cues arent the best for LC so its good practice to test your LC on agar regardless, before use.


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Offlineschpat
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Re: Super cloudy LC. What did I do wrong? [Re: milkboy]
    #28009546 - 10/21/22 03:47 PM (2 years, 2 months ago)

you have way too much sugar in there with the honey and corn syrup.  The LC training camp thread used 0.05% lme (so for your 600ml that would be 0.3g). It's a great thread https://www.shroomery.org/forums/showflat.php/Number/27007467


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Re: Super cloudy LC. What did I do wrong? [Re: schpat]
    #28009728 - 10/21/22 05:46 PM (2 years, 2 months ago)

Is there any discernable advantage to high nutrient LCs whatsoever?
seems like the hard tendency is the opposite in most advice
and discussion


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Invisiblebakedbeings
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Re: Super cloudy LC. What did I do wrong? [Re: smalltalk_canceled] * 1
    #28009730 - 10/21/22 05:46 PM (2 years, 2 months ago)

i much prefer low nute LCs


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Re: Super cloudy LC. What did I do wrong? [Re: bakedbeings]
    #28009736 - 10/21/22 05:53 PM (2 years, 2 months ago)

yeah, you prefer it, but I was in the situation asking the question
if theres any use or specific quirk of high nutrient LC

ive completely moved away from anything above almost nothing for my agar work.. so why wouldnt this work for LCs? dat expansive, rhizo, seeking,
movement.

but is there any use at all for a LC with high nutrtion,
even theoretically

or can we close this avenue, those of us who look into the question,
that high nutrition sucks balls and has no use for LCs and almost
no other use for temporary agar work (most of it) anyway


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: smalltalk_canceled]
    #28009817 - 10/21/22 06:51 PM (2 years, 2 months ago)

This is a good question. The answer would help me a lot.

I've been basing most of what I do off PGT's YouTube channel. His LC recipe is 18 grams corn syrup, I believe 1.2 grams LME, and 1 gram peptone.

He has some incredible results. Isn't this high nutrients per 500 ml water?


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OfflinePadaone
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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555] * 1
    #28009820 - 10/21/22 06:54 PM (2 years, 2 months ago)

PGT...there is your problem. You literally dont need anything more than water and LME. Muy complicado.

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Re: Super cloudy LC. What did I do wrong? [Re: No-Pilot-555] * 1
    #28009822 - 10/21/22 06:55 PM (2 years, 2 months ago)

how dare you link youtube on shroomery, guards!

guards!



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Edited by smalltalk_canceled (10/21/22 06:55 PM)

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Re: Super cloudy LC. What did I do wrong? [Re: smalltalk_canceled]
    #28009997 - 10/21/22 09:45 PM (2 years, 2 months ago)

So, I looked up the dudes video (I assume this is based on his "Super" liquid culture recipe). 12ml corn syrup in 600ml water (or 20ml in a liter) is a 2% v/v solution, that's nothing out of the ordinary. The 1.2g of malt and 1 gram of peptone are .12% and .1% (w/v) respectively, also not unusual.

So, yeah, his recipe isn't really super incredibly high nutrient. That said, it's still overkill for what most amateur mycologists need unless you're getting into really niche stuff. Yeah you're adding a bit of nitrogen and some micronutrients in the mix, but, general saprobes aren't that picky in the short term.

In your particular case, it's almost assuredly the yeast that is causing the cloudiness. Maybe your malt extract was a bit starchy, or you caramelized things slightly, but whole yeast isn't exactly soluble, so I'd point there first.


It's not really going to hurt anything, any fungi that go to town in there will eventually eat up that excess chitin in the mix. It may not look as pretty, and the cloudiness might make spotting contaminants a wee bit trickier, but that's about the worst of it. Also, your mixes are at 3.33% sugar for the honey, and 3% for the corn syrup, which is starting to push things a bit (I usually stick around 1-1.5% myself) but you're not really in a problem range yet (again, unless you're looking at niche stuff). Just a little overkill, but not catastrophically. 


(and really, malt extact on its own is plenty for general saprobes. Maybe you'll have to wait a day or two longer to get to a certain cell density in the mix compared to the video's recipe, but no need to stress about it)


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OfflineNo-Pilot-555
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Re: Super cloudy LC. What did I do wrong? [Re: matsc]
    #28010329 - 10/22/22 07:34 AM (2 years, 2 months ago)

Thank you for this. I've just learned a lot from PGT but in my opinion I like Gary from fresh from the farm fungi. He's more sanitary about his stuff.

Either way, any time I have used what I've learned in my own lab, I tend to end up messing it up a long the way.

I do think my problem is the yeast and I will be getting peptone this upcoming Wednesday so, I'll cut out the yeast and try lowering the other nutrients a bit more.

Thank you for the help! Shroomery is literally the best source for mycology. I love it.


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