Long-term culture storage. Slants, water vials, and centrifuge tubes. With collateral spawn making.

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[@nicohelpdesk435]

I like these long formats, it makes for more details and nice pace. If people find it too long they can watch it in 2x speed or something. Thanks for sharing your knowledge and experience with us. Greatly appreciate.

[@TaylorTEK]

Yes the longer formats 20-30 mins is perfect.

[@TheDarkArtist66]

Thanks for the video ed, this was what i was asking you about a couple weeks ago. Amd the longer videos are definitely better.

[@luismigelgb]

Thanks for this video! I was looking for simple ways to store mycelium long term and I think this is my favorite approach so far. I'm definitely giving this a try.

[@official-kin]

Quality content. Helping me a Lot. Thx for Sharing ur knowledge! ✌️

[@mushroomwonderland1]

I like your channel, good stuff🤙💯

[@chemd1767]

Man's got a shirt on... I don't know if I believe he's ready Ed

[@edwardgrand]

I didn't have pants on ;)

[@andrewbyron9903]

I've learned SO much from watching your videos. Thank you for sharing. 🙏🏽

[@mycomomjeans]

I just learned a ton, thank you. Found you through Mycogeeky, the way you breakdown the "how" while explaining the "why" is just fantastic.

[@psilifunguy]

Hey I appreciate you showing me to cover the labels because the alcohol always erases mine

[@OGTYone]

I’m always self conscious and feel like a failure and I’m messing everything up until I watch you with all your oops and oh craps and you’re amazing and successful at this and it gives me confidence.

[@edwardgrand]

You should try doing it with a time limit, no script, lurking maid, broken air conditioner, and a camera pointed at you. 😆

[@OGTYone]

@@edwardgrand haha but uh no thanks. It’s just awesome I’m super ocd about being sterile. My actual work or job I deal with a lot of sterile environment and cross contamination stuff. When I am working on my myco stuff I alot if the times have oops moments and just seeing you fumbling around and oppsing is a big comfort I just always assume someone like you with as advanced as you are everything has to go perfect and smooth. Buttttt you aren’t perfect and smooth and you know what your doing so it’s reassuring. Appreciate the realness and love the channel. I do have a quick question tho. Is there much a a difference with 1 to 1 shoeboxes and mixes with more sub than spawn I’ve been doing 1/1 and my friend was telling me do 2/1 because 1/1 is more just for genetic work. Other than like 2nd flush does 1/1 effect the first flush in time or yield in your experience.

[@edwardgrand]

​@@OGTYone I'm still experimenting with the spawn/substrate ratio. I haven't really noticed much of a pattern. The hydration of the spawn, substrate recipe/hydrations, actual ingredients (i.e. manufacturer, source), cultigen, age of spawn, type of spawn/grain/sawdust/straw/dung, fruiting container, ambient conditions, misting regimen, temperature inside tub, 'monoculture'/MS/clone/F1 cross, F2,... too many variables for me to make any conclusions. You'd also need to have exactly the same fruiting conditions and a control tub to make any conclusions about changes.

[@nerdnature9592]

Thank you for pointing me to this video Ed. This was exactly what I was needing to know. 🔥

[@edwardgrand]

Glad you found it helpful. There are a lot of little tidbits in the videos, but the title can only be 100 letters long :)

[@TaylorTEK]

You have definitely mastered the art of lighting your alcohol lamp haha man that's ninja quick 🤣

[@edwardgrand]

Lots and lots of practice :)

[@donniebrasco11]

Ok so let's talk about gloves 🧤

[@TheMad106]

Informative video as usual! 👌 I have been actually using these 50 ml tubes to store LCs. Idk if this of a no no but I: 1) Drill two holes with tiniest drill bit I got on the cap 2) Tape one hole with a tiny piece of tyvek envelope and micropore tape 3) Expand the other hole with 1/4” bit and install a rubber injection port 4) Add liquid medium, cap and tape with micropore all around 5) PC for 25 mins and inject with source lc I bought 25 tubes in a rack and they take same space as 4 pint jars. I double bag the whole thing to prevent fridge contamination

[@roycerios8535]

Your videos Rock my dude we totally appreciate you sharing with us. 🍓

[@aidennymes6335]

I dont mind the longer videos. actually i prefer a 20 min video oder a 5 min video. thanks brother

[@depraved420]

Definitely gonna try the water storage, got a bunch of shit on plates I'm tired of transferring lol

[@edwardgrand]

It works brilliantly. All the major culture collections use it.

[@robertosanchez5914]

Thank you for making this.

[@Snufulufugus]

Awesome content! I just did some long term water storage! I found some very nice 15ml glass tubes with the metal lids. Just had to get a silicon cutting mat and a 3/4in hole punch and made some PC safe gaskets.

[@TheTubejunky]

You can use RTV gasket maker. Fill 1/2 inch pvc pipe with it to make a RTV cylinder. Let it dry then press it out and cut it in the thickness you need. Sort of like a lunch meat slicer. Or you could get a two sheets of glass (picture frame glass) and press some in between them to make sheets of RTV. Just remember to use some type of non stick oil or powder to easily remove the sheets.

[@YouLeeBG]

Love your videos Ed ❤

[@patnowhere]

Love it thank you for this!

[@Steveshappylittletrees]

Excellent content 🙂

[@jessembs]

THANK YOU

[@SouthernAsian]

Hey friend, great video. You mentioned the water being able to store for a substantial amount of time but how about the Slants, how long can they be kept?

[@georgecampos7119]

Do you know why the colored agar turns clear? I did some plates and they were blue and some turned clear and it looks like mycelium stop growing. Any ideas

[@SachaLotus]

I have that same question. Idk whether it is contaminated mycelium (cause even after a couple transfers it still happens) or it’s just the mycelium eating the food coloring as well

[@micahgreenlee6694]

I love your content. But I was wondering what size needles do you recommend ??

[@edwardgrand]

18 gauge

[@micahgreenlee6694]

@edwardgrand I was just watching the podcast cast with geeky and he was talking about a 16 gauge. But cool cuzz I already got the 18s thanks for the response 👌 👍

[@edwardgrand]

@@micahgreenlee6694 Either one will work. The difference is minimal. The larger bore (smaller gauge #) just makes it easier to pull/aspirate the LC.

[@CliffordManthey]

Edward where did you get the Freestanding Centrifuge Tubes and The Petri Dish's? Thanks, Cliff

[@edwardgrand]

I bought mine locally, but here is an Amazon link to what I was referring to: www.amazon.com/Conical-Centrifuge-Skirt-Self-Standing-12000xg/dp/B07WRMP7Y7

[@adamschaefer423]

I just bought 100 15ml centrifuge tubes. Can i turn them into water vials since i have a stand for them? And if so how much sterile water would i put in a 15 ml centrifuge tube?

[@TaylorTEK]

Question, why sterile water with no nutrients? I'm just curious why that would last longer than having food for the mycelium maybe less food for contaminants?

[@edwardgrand]

The mushrooms go into status when deprived of oxygen and nutrition. Like a bear hibernating. They slow down their metabolism and just go dormant. I have used this technique for decades and commercial culture collections have used it for over 70 years. They used to overlay agar slants with mineral oil to achieve the same affect, but that is incredibly messy to deal with. I've started to use 50 ml PP5 centrifuge tubes (check online) that have become quite cheap these days. Cultures prepared properly, left in the dark, and not disturbed will last for many, many years using this technique.

[@kingmetroplex]

The information here is great, thank you. How many ml of water do you suggest in the centrifuge?..also I thought you may have been in Australia with all the stuff you did in reverse lol..cheers mate

[@edwardgrand]

I usually use 15 ml in a 50 ml tube. The video thing started as an accident and my ignorance. Now I just think it's funny. Hides my true idenrity. 😆

[@cruzmorgan9358]

Same with monokaryons for long term storage?

[@user-xb1ht4py2v]

What part of Thailand are you?

[@edwardgrand]

20 miles outside of Bangkok.

[@user-xb1ht4py2v]

@@edwardgrand I lived in Bang Pli for 9 years. Wife still in PLuak Dehng.

[@yashuu-rk4lc]

Hloo sir, i have a small doubt can i pasturize my poo substrate in a AUTOCALVE (vertical autoclave ) which has a temprature controller only not pressure controller?? If yes how should I....

[@edwardgrand]

Any type of machine that hits 121 Celsius can be used. Water's boiling point and pressure are related. If the inside of the machine hits 121 C, it means the pressure is about 15 PSI. Autoclave, pressure cooker, sterilizer...vertical, horizontal...they're basically the same thing. Just different pressures and temps. Some of small differences, but much longer story.

[@TaylorTEK]

is that bleach water you have the scalpels in?

[@edwardgrand]

70% denatured lab-grade ethanol (cheaper than isopropanol where I live)

[@the-visual-eye]

Are monokaryons able to be stored in the sterile water for long term or are they something they needs to be used when they are the most viable and fresh

[@edwardgrand]

They can be stored and subcultured the same as dikaryons.

[@maybebanana749]

What do you think of sterilized syringes shot into agar plates to break up myc and suck back up, then deposit into vials?

[@edwardgrand]

Just get syringe caps and leave it in the syringe :)

[@maybebanana749]

@@edwardgrand Thanks for the tip! you are the master.... caught you on sages live last night, awesome man. I read somewhere that one reason the myc lasts so long in water suspension is that there is no fae or nutrient. do you think the agar will shorten its life in suspension? TIA

[@edwardgrand]

@@maybebanana749 The amount of nutrients in the little piece of agar should have minimal effect after it diffuses into the relatively large amount of water. The water will have very little dissolved oxygen because of the sterilization process. I would not suggested shaking it or anything after the chunks are placed inside. The small amount of oxygen above the liquid should be inconsequential. Thanks for watching :)

[@Sage_Advice]

Isn't the 70% iso a fire hazard? I've always been reluctant to have it near an open flame.

[@edwardgrand]

It is good to have something that will cover the container and choke off the oxygen. The lid for the alcohol jar is always nearby. I also have a large beaker that can completely cover the alcohol burner. Alcohol fires will happen. Be ready to extinguish them quickly. Cutting off the oxygen works very well.

[@nicksmith7947]

What do you wrap your plates with? Seems to go a lot faster than the parafilm I’ve been using.

[@edwardgrand]

Grafting tape or food cling film cut into rings. I have another video about it.

[@psilifunguy]

Is that blood agar, and what is blood agar for?? Gd afternoon 4:00 am here LoL 😂

[@edwardgrand]

It's just MEYA with food coloring. I'm on the other side of the world. Blood agar is for growing pathogenic human bacteria.