Tissue culture of cannabis A-Z

Alright folks, it’s time we get a A-Z for tissue culture compiled for the masses of our community that are interested! So with that being said, I’ve been lookin into it myself as well & have only gotten so far with what I’ve found for reading. I know of course it’s a sterilized agave solution with nutes & certain inputs to keep growth either dormant or explode & multiply the cloning abilities. Who’s go the tek? Who’s got recipes? Who’s got stories? Let’s hear it all & get it all laid out so we can level up the community! :call_me_hand:t3::call_me_hand:t3:

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The technique and hormones u need vary from strain to strain, i tried but was not clean enough. Flow hood and pressure cooker are necesarry.
Id practice duplicating bacteria or fungi on petri dishes before jumping into tissue culturing cannabis, once u get a good success rate and hardly any contams, move onto TC.

TC opens a whole new world to propogation, u can even make synthetic seeds from a tissue sample.
Or formulate your homones so that the plants all grow uniform, its wild to see a whole garden of the exaxt same looking plant, nodes and splits are all in the same spot because they had it on a hormone schedule.

I imagine once amazon gets their foot in the door theyll have the whole thing automated from the tc to the grow. All sterile environment and all robotic hands doing all the work

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What nutrients did you use if you remember?

I agree with this, and I also would suggest starting with something that is easier to TC. I said this in the other thread but the book “plants from test tubes” has everything you need to get started. I don’t remember the example they use but they walk you through, step by step, how to propagate a common house plant. Start there then move up.

The biggest bitch is definitely keeping things clean. I haven’t had much contamination but have a fume hood, autoclave and lots of experience growing mushrooms which uses the same aseptic techniques. I’m currently trying to figure out the synthetic seeds thing, but so far I have not had much luck. Have got some to be successful right off the bat but after any storage period germination falls off a cliff.

I got some new strains I am going to put into TC I’ll snap some pics and post some tips if I remember. Going to be a week or two before I get to it

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Curious what others thoughts on the matter are, where do you see the business opportunities with tissue culture? I have a few ideas:

Synthetic seeds: sell to consumers/ business as clean, stable genetics. All growers know how annoying pheno hunting can be, and a lot of people don’t have access to clones because of the legality. If you could sell “seeds” that are true to type it could be huge

Plant preservation: hold and maintain genetics for other breeders and growers so they don’t have to worry about losing genetics to pest, fire, etc…

Consulting: help build and design labs, sell SOPs, etc…

Consumer goods: follow the same model a lot of mushroom companies do where they will sell pre made spawn bags, slants, liquid cultures, things like that but more TC focused.

Breeding: you can essentially “fuse” 2 cells from different plants together to creat a hybrid. If you were to combine this with genetic tracking you could, in theory, test thousands of crosses per week and be much more efficient than typical methods.

All of these seem to be underserved areas of the industry and I’d love to know what y’all have seen or think!

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Assuming these aren’t hazardous chemicals you guys are handling then you could probably cut corners on a fume hood. I normally would never recommend that but assuming the goal here is just sterility and not ventilation of fumes then you could probably build your own.

There are some cheap DIY “glove box” build outs on Google that would probably do the trick for a small sterile work space. Pretty sure they’re basically just big rubbermade bins cut to fit huge dishwasher glove inserts to work through. Presumably with a HEPA filter to let clean air in and a 4” computer fan exhaust. Seal cutouts with caulking or something similar. Pretty sure you could assemble one for like a hundred bucks in materials.

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The synthetic seeds dont have a shelf life, u couldnt sell them retail, it would be a made to order type deal.
The only benefit i see to synthetic seeds over TC is getting a really old seed to pop. U can take the tissue from an unpopped 30 year old seed and make a synthetic seed from it, i dont have much experience, but from talking to my buddies, its much easier to get the synthetic seed to pop then it is to tc the tissue from the inside of an old seed directly.

The breeding aspect doesnt work with tc ur thinking of crispr :wink::wink:

The best use for TC is to reinvigorate old diseased clones, and keep a sterile backup in a bank.

One business model i thought of is a clone babysitting scheme, where u send in ur plant u want TC, u pay for a storage and maintenance fee and at any time u pay a flat rate to have a clone sent back to u.
A clone bank basically.

All clones are sent and kept under an encrypted directory, so only u know whats in ur storage, each clone u send in has a unique ID code and only u have access to that ID. Stops people from robbing each other clones

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You are right, that is crispr but it would still be super cool! Just need to go get a PhD in plant genetics real quick before I pull the trigger on that one :laughing:

I think you could make a model out of the synthetic seeds though, and they do have a limited shelf life. Some plants have 80%+ germ rate after 6 months of cryogenic storage so there would definitely be a way to make that work (assuming you can do that with canna). I think the biggest advantage of the synseeds over TC is the fact you can propagate them ex vitro, which gives the layman a good chance of success with them.

I do like the idea of the clone babysitting, but I think the hardest part of that would be convincing people you aren’t going to Jack their genetics and sell them elsewhere. With the state of the industry, that would be a fair fear to have, sadly.

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Thats why its all encrypted, all the workers have no idea what is what. Just codes that look like this ahao763hkkHKGf2h2i

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I got what you’re saying now, that’s a great idea.

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Mirashige and Skoog media with generic agar works just fine. Cannabis is actually fairly forgiving and you can basically rip an SOP for mammalian TC and it’ll be way overkill.

You can make a DIY LF good but honestly, just buy one and be happy, it’s the place to splurge on this project.

As far as business models, there are a few:

Easiest and most important: cryogenic tissue storage - bank em for later like you would with seeds, except you get to know exactly what will pop.

Next: clonal propagation of autoflowering plants. No more culling males from the field.

Moving up to the hard stuff: processing for viral removal without identification (it’s a thing, check it out).

Also: sexual propagation of two female plants. Also a thing called a gene gun which is, amazingly, exactly what it sounds like.

Lastly: every knockout or viral-based genetic modification technique including CRISPR.

Or: just growing a fuckload of explants in a test tube rack which looks really cool

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This is what I like to hear & see! Plenty of info for others to dive into the rabbit hole & get a grasp of what direction to head! Thank you :pray:t3::call_me_hand:t3:

I just googled “plant tissue culture procedure” and there’s like 75 good write ups, most of which are pretty similar. Also, I’m not sure if it’s still out there but I think Fisher has a free interactive mammalian TC lab class which is pretty solid for teaching someone how to work in the hood without being hands on.

Like I said before, 99.9% of the job is keeping stuff clean. You can almost just go through the motions with a piece of dead plant material, and if your agar doesn’t turn into a tube of fuzzy death you’ll pretty much have success (with propagating anyways, I never got near any of these other processes).

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Let’s group fund this for the clone group

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I’ll look in storage I believe we still have a flow hood just collecting dust.

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These are all good ideas, How would you rate the start up cost of each of these ideas, and time required.

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I bought a kit from DIY tissue culture, looks pretty complete. @diy_tissue_culture
i have yet to use it, too many projects going on all the time and i need to set up my hood and clean tent area somewhere in my facility.
cant wait to try it out. will let everyone know the results.

thanks!
tom

Hehe :)) cleanliness of us does very by proffesion :)) I think for a test of the laminar you shoud try to breathe on the agar plate fro a distance of 30-50cm… Just to see if the flow is gonna be saving your ass or not.

If it doesnt turn into CFU in a week on you, your on your way to cleaning your shiny hands in some isopropanol with your tools… 3x times

And then the fun can begin…

Oh right gotto wash the plant, yeah right right…
Ok great lets go…

Now gotto wash the plant…

… Okay Fuuuuck… reached for my dirty phone outside of the hood flow cause my girl texted me…

Back to step one of SOP… ( you said to share a story)

And then… Then this was the fun part:slight_smile:

imma go for it ya’ll got me wound up like a swiss coocko clock…

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