12 replies to this topic

[larryl]

Thanks. I have a few questions from time to time and hope to contribute. I'm working now with a liquid culture for a cubensis variety. I am finding the best results with a plain dextrose 4% solution. This or other simple sugar solutions are what others seem to recommend. 

 

When I add other components like trace minerals, growth seems to slow way down. When I add other components like malt, which I might think would be a more complete nutrient, then I see more contamination. I guess I'm puzzled since glucose is just C, H and O. I would think that some nitrogen would be in order to make proteins. So I guess I'll just go back to dextrose. BTW, I'm inoculating the LC with blended colonized PDA with a mycelial isolate. Perhaps the PDA that is initially chunked into the blender contributes enough nutes to start things off.

 

At one point I used half (75cc) of the LC after 7 days of incubation with a stir bar and I added an extra 75cc of dextrose solution to the LC hoping to restore the LC. It seemed to survive but did not get very dense like the first time. Perhaps nitrogen had been depleted by this time. I plated the LC to check for contams and it looked clean at 4 days with a few mycelial colonies but only about 1/3 as colony dense as the original mature LC. 

 

So I guess my questions are:

 

1) Agreement on dex or other sugar for LC? VS more complex nutrient solutions?

2) Can you feed your LC and keep it going? If so, then with what? Adding 4% dextrose has not seemed to promote much growth if any at all.

 

 

[FunG]

Simple sugars work fine for lc's , dextrose is the only sugar I've used back when I tinkered with liquid cultures so I'm not entirely certain about adding in more complex nutrients like brf and I've only read about others restoring lc's the way you tryed with successful results.

If you feel strongly about nitrogen playing a role in enhancing or promoting growth you could try adding in a couple cc's of coffee....that would add nitrogen and complex nutrients but it may throw the p.h off just slightly.... another mineral to add would be a pinch of gypsum.... that would bolster the mycelium and prevent a p.h swing by adding coffee.

[PJammer24]

karo syrup, dextrose, grain water, malt... These are some of the typical LC components...  There isn't much reason to add anything extra.

 

Don't use BRF or Coffee... You could use the water drained from the BRF or coffee but not BRF or coffee in the raw due to their not being water soluble... This seems like a bad suggestion.

[FunG]

Brf is water soluble

It just turns the water murky

And by coffee I assume they'd know liquid coffee not the grounds.

P.cubensis love there coffee probably due to complex nutrients and high nitrogen, I wouldn't suggest it for other species....he did state that it's for p.cube.

[coorsmikey]

4% is borderline too much. I know its all over the web and shit but that doesn't mean its best, It just means it works. Cut it  down to 2% and then the additional supplements won't slow it down too much. In higher quantities sugars begin to work like a preservative. Ever see a moldy candy bar? 

No need to feed an LC, Make more and take a sample from the old one to the new one. Use it to inoculate grain and cakes to grow mushrooms, not see how long  you can keep alive. Change the nutrients up every LC you transfer to and you can keep a strain alive for years without senescing. Less sugars and nutrients is better than more with LC.

[PJammer24]

Brf is water soluble

It just turns the water murky

And by coffee I assume they'd know liquid coffee not the grounds.

P.cubensis love there coffee probably due to complex nutrients and high nitrogen, I wouldn't suggest it for other species....he did state that it's for p.cube.

 

brf is no more water soluble than any other flour or floured grain... Suspending brf in your LC mix is not the same as it being soluble

 

Considering you can often see a contamination in the LC due to its being foggy, I try to keep my LC as clear as possible... You can often tell a foggy contaminated LC even when the sugars have caramelized. I try to avoid caramelizing the sugars but you can still see the difference between a clear, uncontaminated LC and one that is foggy...

 

Using drained water from hydrating grains, BRF is essentially a floured grain, is a good way to make agar plates but in my opinion is not well suited to LC... LC is super easy using typical approaches with sugars. The water from hydrating grains, which would include BRF, is filled with the starches from the grains but not the grains themselves... While this would likely work for an LC, I don't view it as being the best approach. If you want to make use of the water, use it in agar.

 

Actually suspending BRF in your LC is not something I would suggest. You don't hydrate BRF in the same manner that you do typical grains so you would never have the leftover, starchy water you can collect from other grains...

Edited by PJammer24, 11 March 2020 - 01:07 PM.

[larryl]

 

4% is borderline too much. I know its all over the web and shit but that doesn't mean its best, It just means it works. Cut it  down to 2% and then the additional supplements won't slow it down too much. In higher quantities sugars begin to work like a preservative. Ever see a moldy candy bar? 

No need to feed an LC, Make more and take a sample from the old one to the new one. Use it to inoculate grain and cakes to grow mushrooms, not see how long  you can keep alive. Change the nutrients up every LC you transfer to and you can keep a strain alive for years without senescing. Less sugars and nutrients is better than more with LC.

Thank you, CM. Good to know about 4%. I think I'll settle on 2-3%. 

 

What are thoughts about other nutes like malt extract. Just a thought to add other basic elements besides C,H,O. But I have a hard time arguing with success. 

 

I discovered something else a bit by accident. I added 4% dextrose to a sterile jar that I made for agar work. It had a layer of agar on the bottom. I nuked it to melt the agar and swirled the entire mix until homogenous, then let it cool and added about 10cc of left over cube LC from a prior jar. The final volume is about 200 cc. The mix is a bit viscous from the bit of agar. I'll see how it goes. Perhaps the agar (PDA) will provide some added substrate and some nutes. IDK but might be interesting.

[The_Tripper]

I've had much success with just agave nectar in distilled water, no need to overcomplicate the lc, save nutes and recipe changes for your subs.

[goldenteacher1163]

LC is just the vehicle to give you a nice quick burst of growth and an expansion of useable spore solution. Less seems to be better when adding your sugar source (2% as mentioned above is great). Adding nutes can be counter productive by clouding your LC etc...my friend always tries to use it up as soon as growth starts to really take off (this gives you the hot inoculation that makes LC great) Simple is the way to go

[Tom25]

Just wondering how many people test their lc out on agar just to be certain it’s pure? If so, how many Petri dishes- I’ve heard some people do up to three to be absolutely sure?

[Myc]

Welcome Tom25 !

Testing your liquid cultures on agar is just good laboratory practice.

I usually just use two plates from the same batch. One petri is left blank as a control to make sure the agar is clean. The other gets inoculated with a drop of LC.

[Tom25]

Thanks Myc, great advice!

[Enigkotik]

I used the recipe I got from the psilocybin mushroom bible by Virginia Hayes and Dr k Mandrake. Very simple recipe, 12 g of honey, 288 g of water. I made five at the same time, four of which are PCs, and one lion's mane. They are all doing great.