Liquid culture (LC) technique. Expanding liquid culture, inoculating grain, storage, and discussion.
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- Опубликовано: 28 авг 2024
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Summary of things I’ve been doing wrong: flaming a fresh needle, violently shaking my LC, having a little air in my syringe. 😅 thank for the info ed. it’s truly invaluable
Liquid culture is a great. The first time I got a syringe of culture I thought 'Why can't I make this stuff and use a little bit of the culture I bought to keep it going?' Now I do it every time, I can keep cultures for ages and inoculate grains without having to buy more if anything goes wrong. Cheers dude, love your quick fire videos 👍
I think this is the best liquid culture video I've seen, I like how you've cut through the bulls#!t, nice work Ed.
Really dig the simple approach to LC and bags! 👍 👍
I use the 10 ml vials with liquid culture to store masters in and have been really happy with them. I can do 100s of them at a time in the pc soo it’s nothing to transfer to a new vial once a year if need be.
I started my first spawn bag a week ago and the mycelium is already 15%+ looking beautiful... now i cant stop watching mycology videos 😂
Ive even bought 2 more grain bags, a boomer shroomer tub and her substrate...and a still air box 😭😂
I think i have a new hobby atleast for a little while
I get better with every video you release! Thank you for your insight.
Ive been collecting my grain water from hydrating my popcorn. I add a small amount of peptones and use it for culture and agar. Been working well for me
Love watching your videos learning alot Thanks so much
Your videos are amazing and so thorough. Expert training! Thank you.
Awesome Thankyou I was the one on the other video asking about sealing the tape after inoculating the bags.
Stapling the grow bag is unbelievably convenient
It broke my heart when I had to shred my Lion`s Mane mycelium in the jar. This LC and some failed attempts are my first steps to create fungi-life. I was so happy to see it growing the jar, shredding it AND trying to take it out to inoculate did hurt. It was so peacefully swirling in the liquid...and any disturbance might cause contamination.
No, not how it works buddy. No such thing as spontaneous generation. You need spores or colonies, etc, you need contamination physically to be present and alive. Just one spore, or microscopic amount is all it takes. Shaking does not produce contamination magically, if contamination is present in a single colony than shaking will redistribute that contamination making its presence much more known.
@@TacosGaming you took too much. Not sure who you reply to but I did not mention the word "contamination". My LC was not and is not contaminated. I just did not get mycelium out of the jar, only the liquid. This is what I called a failed attempt. I did not mention "spontaneous generation" either. No idea where you got that from. As said, you take way too much to be with "us".
Thank you so much for this knowledge, Im doing a side by side with all kinds of measurements of everything I have left , then when it’s all gone I will be doing .5% LME only!! Thank you for sharing your knowledge sir.
what is LME?
Would be awesome to see you run grains in the pressure cooker or autoclave. I’m Always picking up on something in your videos. the do and don’t of grains would be awesome. People over complicate everything soo I love when you confirm when something is bs.
There are so many autoclaves, and pressure cookers. I'm not sure where to start.
Yeah I agree, the more video Ed makes the more bs info we can all filter out and this guy can show the basics upto the mad scientist end of the spectrum. I learn every video even if it's subject matter I've watched hundreds of times. Keep it up Ed.
I did my LC 15 mins for ages and I never had any problems. I even had caramelization problems if i did more than 15 mins. I think it really depends on some other factors, especially the pressure. liquids in general are way better in heating up fast and evenly inside a clav.
If you go for .5% sugar in recipe shouldn't have caramalisation issues.
Why do you day problems....caramelized lc does the same as mom caramelized....look up videos on the topic. It does not matter
15mins at 15 psi is no problem. This guy is doing a hour cause he has a 12psi pressure cooker. If you have a 13 psi pressure cooker you do 30mins and 15 psi 15 mins.
"This isn't like a bag of popcorn ..it's literally a bag of popcorn"🤣🤣🤣🤣💀
LC for 30min at 15psi has worked great thus far, but longer isn't gonna hurt.
Thank you so much Ed that answered so many questions
Hey bro, are you using oven bags? Do you have a video of you prepping grain in those bags by chance? Love the videos btw. You inspired me to buy a microscope!
Great video! I love how well you explain. Could you please share how long the mycelium remains viable in the liquid culture jar?
It is deprived of oxygen and goes into hibernation/dormancy. The same thing will happen if you keep your plates sealed and they very slowing dry out. I have revived 6 month old plates. As soon as they hit fresh agar, they usually start regrowing.
Great educational video
RADICAL 👍
Well now I have all this LC. What ever shall I do with it all? Two and a half months no contam. I am a mushie master with all of 6 grows under my belt. Bow down before me. You make it too easy Ed.
Yes. Success can be a bit of a burden and a blessing. Start looking for a new house. It only gets worse. :)
I think I've just watched this again for the like.. 10th time, easy! Something I've always been wondering.. can syringes and/or needles used for LC inoculation be PCed and re used? I'm running through quite a lot, they're cheap but it's a lot of plastic in the bin..
Yes, they can be PCed, but they tend to distort and become opaque.
Hole punch makes quick holes on lids
Dude awesome LC video. How long do you let your LC expand before you store it in refrigeration? I have a few jars that are about 8-9 days in but I’m not sure when to slow it down.
Thanks Ed
Would you suggest Keeping LC in the dark for better colonization
would you explain briefly why the staples in the bag works vs the filter patch? main question is contam - i am very new to all this little over 6 months. i use filter patch bags and have been adjusting my technique after watching your videos but every time i see the folded bag with staples i wonder if thats the best route! thanks for your time :)
I don't know if I'd consider them a replacement for a filter patch, but they work well for me and are easier/cheaper. I think it's about the right ratio of grain to air in the bag. I use about 500g of cooked grain in a 12x18 inch bag. I try to keep the upper half free. You should give it a try if you have access to cheap PP5 bags like I do here in Asia.
when how does temperature play a role with liquid ie do you use climate control thank you for your strait forward videos
Temperature will just affect growth rate. Mine just sits at room temperature. Like 27C.
Hi good Doc! I have an eclectic question that I was hoping you might be able to answer for me! Is there any way to estimate how much mycelium is available in LC? I'm still using both agar and LC to inoculate grain jars, and I want to give each jar approximately the same amount of mycelium but I have no idea how to convert the amount of mycelium available on a plate vs how much is available in a syringe of LC.
Would it be reasonable to assume that 20mL of fully colonized(well shaken up) LC contains roughly the same amount of mycelium as a fully colonized plate (with 20mL of agar) if both were made with 0.5% LME w/v?
Thank u so much for the video.
I was wondering if boiling a potato and using some of its water to add to the LC would work instead of the LME🤔
Yes. It would work well. You'll just have to figure out the concentration, time of cooking, weights, etc.
Hey Ed … where do you get your plastic lids???
We have an online place called Shopee. Sells everything. Just make sure they match the jars. Many sizes.
It looks like a pint-ish jar you are using? Like half full? Is less best as far as total quantity? Could one increase the volume? Get the culture closer to the top? I started some water and honey a couple weeks ago. I forgot the marbles lol it seems strong. I followed fresh from the farm and tested on agar for contamination. About to transfer and do it in lme and yeast, with marbles! Great timing thanks!
That jar is 500 ml. The liquid is about 150 ml. I like to keep it lower in the jar so it doesn't get near the rim when swirling. That is where contamination will sneak in. All LC recipes are fine. Just don't go past 4% sugar and don't contaminate it :)
And don't forget the marbles. LOL
Have you heard of anybody purposely fruiting in LC? I've only accidentally fruited in LC when forgetting to use jars.
Btw I see you're a fan of corn! Is this due to local availability or do you think it's the most ideal grain for cubes? Do you happen to have any idea whether there is any correlation between the protein content of grain and its suitability for cubes? The reason I ask is I notice that nobody uses any high protein grains like quinoa, but seem to prefer moderate-protein grains like brown rice, millet, oats, or the lower protein grains like corn and rye. I've been thinking of using mixtures of grain, but am not sure if including quinoa is wise. Thanks again for fighting the good fight, Doctor!
I've never heard of anyone purposefully doing it. They do fruit if left alone for months.
I am a fan of any grain that is cheap and readily available. I don't think the protein matters. All grains have abundant protein for the production of alkaloids. Mixtures of grains work well, but they need to be prepped separately before going in the PC. I have found corn and rice works very well. Wheat and corn too. But they require very different preps. If I can get one grain cheaply, I just focus on that one and doing it right. That is more important than the nutritional profile.
Thank you for the kind words and watching :)
Silly question, but what kind of fuel is in the lamp that you flame the needle with? I currently use a torch, but I like the idea of a lamp. There is a special name for that lamp, I believe, but I can recall what the name is.
95% denatured ethanol is the fuel. I think it's just called an 'alcohol burner'. You might be thinking Bunsen burner. That is too hot...and expensive. You can DIY an alcohol burner. Check RUclips 😉
ruclips.net/video/65HTaBhPjLQ/видео.html
Thank you!
Hey Edward. How long does it usually take for spores to germinate in lc broth? 1 to 3 weeks?
Depends on the age. If nothing is going on a 3 weeks, it'd be suspicious. I wouldn't use spores directly to LC though. Very likely to contaminate.
@edwardgrand I did it around a week ago. It looks like there are tiny specs of mycelium in. Very tiny, tho. I know it's likely to contam, but I'm hoping it works out. Put some from the same spore syringe on agar, and nothing has shown up yet. Strain is APE.
@@devilsadvocate1338 APE syringe is probably LC already. Albinos don't drop spores, so it is very difficult to make a spore syringe.
@@edwardgrand oh damn! So that's even better!
Thank you very much for all of your valuable info in your videos.
Quick question, you ever tried erithrytol LC? I'd assume it'd work fine too and less prone to contam than mea somehow?
Never tried. Contamination comes from technique, not ingredients. If the recipe will grow mycelium, it will grow contaminants just as readily.
Great video! How many times you can make LC from LC before you cultures start to die?
I haven't really noticed any degradation after 3 subcultures. I usually clone a good fruit and grow it out on agar, then use that to inoculate a new jar of LC. I also go back to spores frequently.
@@edwardgrand When you say frequently do you mean every year or?
@@moviesandseriesrecap 6 months
@@edwardgrand THX!
How are the staples which are not sterile not contaminating the bag by puncturing it?
You have to make sure the staples only go through the folded parts. I make sure the fold is about 1/4 inch wide and staple as near to the middle as possible. Look closely where the staples are actually going through the bag.
Hey I've got a question about LC. I run into an issue where I think I'm over twirling (or magnetic stirring, I only have 10 stir bars so I use them if they're available) and the blessed myc will get more thick and less whispy, and starts to sit at the bottom & is no longer useful- I've put it on agar and had it come to life 2 WEEKS LATER, but usually it does not revive. Is there a trick to bring it back and I'm assuming the issue Is over agitating, do you think that could be the culprit?
Also, how long after innoculating w/ agar until you are ready to use the LC? How long until it's technically usable and how long until it's comfortably usable w/ confidence? Thanks man, you have been invaluable to this process as I transition from "enthusiast" to "Balls Deep" into Mycology.
Too little agitation and the myc will run out of oxygen and stagnate/die. Too much and I think it will overgrow and use all the nutrients. The waste products might build up and cause weak grow and recovery. It is a fine balance. There are so many variables to consider to make general guidelines. I usually try to get the myc to optimum density in about 2 weeks. I agitate once or twice a days for about 5 minutes. Once it hits that optimal density, I would use it, pull it and refrigerate it, or just let it sit. If the LC just goes dormant (because of low oxygen), but doesn't die, it will revive when oxygen/agitation is reintroduced or it is put on/in fresh media/LC. If you don't get around to using it or pulling it into syringes (and refrigerating), I'd just let it sit quietly until you're ready to proceed. I have restarted 2 year-old LC that was just sitting quietly at the back of a shelf. As long as it stays clean, no problem.
@@edwardgrand right on, thanks, I do notice that my lc is losing its amber color in those jars - so i suppose its over consumed the nutrients do to my excessive agitation? i would not have thought of that, i was like "damn, i beat that myc up?"
You recap and save your LC syringe. What are your thoughts on boiling and reusing the syringe and needle on a totally different LC? I've come across advice saying 15minutes in boiling water is OK. My guess is they should really be PC'd.
I wouldn't reuse them. The PC distorts the barrel and plunger. They are cheap. Not worth the time to reuse.
❤❤
Hey brother, what kind of bags are you using for your corn there. I'm in a search for a bagging system and not having any luck. I wanted to use food saver bags, but according to the internet experts, that is a bad idea.
They have to be PP% (polypropylene recycle code 5) in order to hold up in the pressure cooker. Any other plastics will melt or get heavily distorted.
i got you a nice project if no one has done it . fill a qt mason jar with pieces of agar and use that as grain and see what happens . let me know what you think and if someone has done it
No one would do that.
If you do a needle biopsy that’s essentially a clone to LC?
yes
@@edwardgrand ahh ok
@@edwardgrand I’ve been looking online for this answer and I can’t find it so I figured I would ask you! Are purple mystics coir or dung loving mushrooms?
coir@@Houseofmycology
@@edwardgrand sweet I got some from a field and I have been germinating the spores on agar and I have some colonizing on popcorn! But didn’t know if I needed dung or not! Thanks brother
What if you are running low on lc , can you just add more sugar water to it . I know it’ll be a 3 gen then but would it work ?
It would be better/easier to make a new container of LC media and inoculate it with a bit of the remaining LC. Using the old bottle and adding stuff to it sounds problematic.
Why a 18 gauge needle? Why not a bigger one? 16? 12? How does koolaid work as a LC?
18 gauge is easily available and cheap. Kool-aid doesn't work for LC.
What kind of flow hood are you using? It is super professional and I want one.
Not sure the model or manufacturer.
Yo, i have a question. If i cover my lids with foil and after pcing i leave lc inside to cool overnight, can i then remove a foil inside sub and not sanitize the port
Inside the SAB? I guess you wouldn't need to, but alcohol is cheap.
@@edwardgrand thanks for the answer i usually use bleach 1:10 is it good?
Hlo sir, i have a vertical autoclave which temperature adjustable so can I pasturize my poo substrate, if yes how do I need to proceed ??
Just operate it as normal. Depending on the volume and type of substrate, you may need to adjust the temperature and time.
@@edwardgrand sir, so can I pasturize my poo substrate at 5psi (75'c) ??
@@yashuu-rk4lc 5 psi will require like 8 hours to sterilize. Don't know what level of sterility you want.
@@edwardgrand nah sir, i want to pasturize my substrate ( killing the bad bacteria and leaving the good bacteria), am not talking abt the sterilization ( killing every micro org)
@yashuu1396 An autoclave is for sterilizing at higher pressures. Pasteurization should be done at atmospheric pressure.
What temp range are your LC jars kept at?
Fahrenheit please...lol
Normal human temperatures. I don't do anything special.
@@edwardgrand so room temp and not refrigerated?
As always, thanks for wearing clothes 😂.
I'm naked from the waist down...
Mushrooms have survived 800 million years without human intervention or assistance so all this human silliness about sterilisation makes me laugh
We are trying to grow a single species. We need to eliminate the competition or replicate the exact conditions that have been fine-tuned by mother nature for 800 million years. Since we've only been around for a tiny blip of that time, we have to make due with our current knowledge of how to encourage them. If you want random mushrooms to pop-up, just let mother nature take over. She does fine by herself.
You obviously don’t have a very good understanding of cultivating mushrooms in a controlled environment if that’s your perspective. In nature, billions of spores are released and only a very tiny percentage of those will germinate and produce mycelium and then grow into mushrooms. In the wild they are competing with millions of other organisms, bacteria and different fungi for a finite amount of resources in any given area. When cultivating at home or in a lab, you sterilize to eliminate as much competition as possible to give your mycelium the best chance of success in ideal conditions . Now, if you want to try the natural way and just throw your spores into some dirt and hope they produce mushrooms without human interference, go ahead and let us know how that works out. There’s a reason everybody talks about sterilization, airflow and temperature . So keep on laughing without an actual understanding of what’s going on. As they say, ignorance is bliss.
These are the types of comments you get when people are ignorant.
“This is not a bag of popcorn” while holding a bag of popcorn