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Tissue Cultured Marijuana ? You Bet !


trichcycler

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After I decide on a recipe for my next culture it's time to mix. I measure and mix a liter at a time, filling around 30 baby food jars(bfj) or many more test tubes. My glass and tubes are already cleaned and have been boiled too. Not sterile, just clean. In a one liter measuring cup I add water, hormone(s), sugar, agar(the stuff they make bite molds with), and nutrients. The ratios are dependent upon expected outcome. Some may be intended for rooting, and some only sprouting, without roots, some for short stout plants(think bonsai) or some for freak shows(pics coming).

I use a magnetic stirrer to keep the mix in solution 41WDJB5HJfL._AA160_.jpg

The ingredients are sometimes difficult to dissolve, and this is a hand saver. I can perform other duties while it works silently.

After mixed I 1/3 fill bbj and test tubes as needed, and place caps lightly on them. The caps are loose to allow hot air to escape, but tight enough to keep water from splashing in. I stack them in a pressure cooker neatly. If I have room I include tools like my scalpels, some paper towels, a mason jar of water etc. These always come in handy in the culture lab(tent).  Sterile water to work with is a must. If you use spring water as I do you'll notice the minerals separating from the water after its sterilized under pressure, no worries though, the water is drawn up via syringe, baster, or eye dropper, and no chunks get in.

I keep a stock of sealed sterile jars,tubes, tools at the ready for the next excisions.

 

It sounds silly, but this sonic toothbrush cleaner has been a turning point for my mold free successes 419QKnNcNxL._AA160_.jpg

IN the plant cleaning process alcohol, bleach and water are used in separate steps to remove any pathogens, mold, bacteria etc. The sonic action of this cheap thing is unmatched cleaning power I've never had shaking samples in a jar of liquid. You'd think bubbling away in bleach, then water, then iso alcohol, then bleach then water again would kill plant material, and it does. The damaged bits are trimmed off before planting the explant sample. Callus material is grown, as undifferentiated plant dna. It is no longer a cannabis plant, but a blob of dna awaiting a natural hormone trigger to respond as ordered.

 

 

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I haven't spent the time to go through your links yet, but I intend to. This has got to be one of the most interesting topics that I've seen in a long time. I'm really grateful you're sharing insight.  

 

Please tell me that you've played with this:

 

Propagation through alginate encapsulation of axillary buds of Cannabis sativa L.—an important medicinal plant

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I call those my artificial seeds. They are precisely what is discussed in the link you shared. thank you. I don't like using antibiotics in culture, and some of the other hormones for this type of culturing, but I have, and occasionally do. It requires some real care to avoid exposure. After starting with crushed vitamins, sugar, jello, basal salts, and eventually graduating to a full lab of all the goodies, you'd be surprised to know I've devolved, and attempt to culture in a more natural way since growing cannabis. Seemed I cared less with flowers, but cannabis changed my mind. My Willow Water is my friend.

 

I'd like to add that when mold happens in a jar, and it does, the jar should be boiled before disposing the contents. Pouring them down the drain is not an option. I've cultured plant cells, mycelium ,animal cells, bacteria,insect cells, ecoli,(for jellyfish protein feed) and human cells. The molds that are produced are some of the most far out stuff I've seen.  The wildest of all comes from contaminated human cultures. some of these are dangerous, don't do them until you know how to protect your surroundings. Most infected cultures I've seen end up being an un sterilized human cell/hair, etc  dropped into the jar before sealing, during transfer. I bet they are nasty to come in contact with.

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big fun happens when taking explants from different parts of the cannabis plant for culturing. For instance the arguably easiest to culture is a meristem end. But every part of the plant can be cultured, from a root bit, to anthers, to seeds, calyx, stick, leaf. each may result in a different type of arrangement. Some are real freaks, especially if you get the hormones wrong. These are no exception

 

 

 

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auxins, cytokinins, gibberellins, abscisic acid, ethylene Usually used in conjuction. They are naturally occurring growth regulators, even in humans. A typical mix might look like (for instance) one drop of an auxin, mixed with 1 gallon of water. With only a drop of that mix, mixed into a liter of pure water, the (.00000) is amount used is very small, but very effective. There are a couple dozen of each used in culture, however I only used a few,naa,iaa,iba, zeatin, bap,These hormones are exactly what cannabis uses to communicate with itself to, increase internode spacing, produce more root structures, grow taller, or bushier, stop or begin producing flower sets, etc.

Today I use aspirin, willow water instead. a longer process, but no need to store these chems anymore. 

 

Topping the cannabis plant sends hormonal signals down the plant structure and causes it to produce more lower structure and bush out. The same can be seen with a lollipop trim habit, causing a plant to reach up farther. these are most likely a response to an attack, and the need to survive to make seeds or to fertilize other females.

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How was that bottom one? man I wish I wasnt fricking getting old and blind lmao!  How did the bottom one turn out as far as potency/taiste?

 

This is the most interesting thread ive ever read any where!!  What else can you put out there?  Man I would love to pick your brain, when god said line up for brains, I thought he said trains! lmao

 

Peace

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she was a cultured Iranian fast flower grown outdoors. other cultured ones indoors showed the same freak, but the normal clones did not. dr greenthumb has these, hashy, stony effect, huge harvest weights. normally I root rather chute in culture, but this batch of Iranians decided to chute instead of root. wasn't sure if this was my error in mixing or what, so I paid attention and got freaks. I don't grow this strain anymore, but its the biggest producing strain I ever grew, straight from seed .

 

the guitar one is a blzbud, upper right corner of tent pic. she vegged the same time and same nutes as the others around her. those were from clones, but she was a cultured error. they all were awesome though in the end, and the freak gave up big buds too.

the right side is blzbud

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I've revegged a couple times just to see it work, but I found it troublesome in hydro and in dirt so dismissed it yrs ago. I always harvest whole plants at once. I grow them small generally and expect only a couple ounces each. that affords me less work, and a focus on variety rather than weight. When they're ready, they're ready, I wouldn't let them ripen one more moment.

That bottom pic looks just like you picked some buds and let it grow some more at 12/12. The top pic looks like a re-veg.

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  • 3 weeks later...

Take a vacation and see! lol,  One of these days we will have it as good or better than CO, we just need a few more yrs and to get these old phaqs out of politics, than the people who have used mj all of their life will be in office!

 

I mean we went from I didnt in hale, to I inhaled and tryed coke, to I inhaled and done lots of schtuff!

 

I totaly beleive some one with your knowledge will do great in this state, no matter how they lay out the grow rights when mm supply stores get legal and certain growers get legal, they will all be interested in what you have!

 

Hang in there, keep doing what you do!  I dont get mm from you but I most def apreciate your knowledge!

 

Peace

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What would a bio luminescent cannabis plant be worth to a Dispensary in Colorado ?

offspring phenotypes and clones/cultures would be capable of the  expression .

You know, I think if I were to see a grow with a bunch of bright little neon ladies in the dark I'd be pretty ecstatic. That'd just be the bee's knee's in my book lol.

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whadda guy, thanks for he kind words, you phaquing phaq !! :hair:  :blow-a-heart:

Take a vacation and see! lol,  One of these days we will have it as good or better than CO, we just need a few more yrs and to get these old phaqs out of politics, than the people who have used mj all of their life will be in office!

 

I mean we went from I didnt in hale, to I inhaled and tryed coke, to I inhaled and done lots of schtuff!

 

I totaly beleive some one with your knowledge will do great in this state, no matter how they lay out the grow rights when mm supply stores get legal and certain growers get legal, they will all be interested in what you have!

 

Hang in there, keep doing what you do!  I dont get mm from you but I most def apreciate your knowledge!

 

Peace

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I have a santa marta strain in culture and she is an auto. One jar of material can produce over a thousand clones per month, or alternatively a jar of roots can yield twice that. the auto's are identical in culture, until they chute. The regs just keep vegging, and the auto's go into flower regardless of light, just like normal. I did it because that was the best auto I've sampled, and just like you was curious. I really have no use for an auto, however, I am fascinated by them, and think we'll here a lot more about them for their high cbd accounts. sorry, beat ya to it....don't let that stop you though

Also, been going through the articles: I wonder if itd be possible to clone auto's more successfully by using tissue culturing....Oh man, I've got to try that sometime if you don't get to it first. 

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I have a santa marta strain in culture and she is an auto. One jar of material can produce over a thousand clones per month, or alternatively a jar of roots can yield twice that. the auto's are identical in culture, until they chute. The regs just keep vegging, and the auto's go into flower regardless of light, just like normal. I did it because that was the best auto I've sampled, and just like you was curious. I really have no use for an auto, however, I am fascinated by them, and think we'll here a lot more about them for their high cbd accounts. sorry, beat ya to it....don't let that stop you though

I've got to learn how to do this. Really.

 

Point me into the direction of an introductory kit or childs play set so that I can move that ball. I'm new, but I love the whole learning process thing and this seems like some really good future proofing stuff to ahead of the game on.

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google kitchen culture kits, you are on your way. buy "Plants From Test Tubes" and your will be talking soon !

I've got to learn how to do this. Really.

 

Point me into the direction of an introductory kit or childs play set so that I can move that ball. I'm new, but I love the whole learning process thing and this seems like some really good future proofing stuff to ahead of the game on.

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  • 6 months later...

I've got to learn how to do this. Really.

 

Point me into the direction of an introductory kit or childs play set so that I can move that ball. I'm new, but I love the whole learning process thing and this seems like some really good future proofing stuff to ahead of the game on.

You're already successfully cloning, which puts you about half way there! Sterile technique is a bitich for some, but honestly I followed directions once, and have no persistent issues as a result. In my early days I utilized regular cloning powders(diluted to near nothing though) in my rudimentary attempts through to successful ones. follow the direction for culturing carrots was the ticket to quick results, trials, experiments, etc. From those I learned to actually bring these little gals to small trees over and over.

Skip the flow hood, unless you're rich I suppose. I skipped fans, exhaust, air/heap filtration and spent money instead on a small grow tent, Lysol, bleach, pressure cooker, alcohol burners, mason jars, babay food jars, sonic toothbrush cleaner, hormones, books. with a little luck and a year to play, maybe a couple hours a week, a beginner with no knowledge can culture from a living mj plant, establish, multiply/divide, root, chute with little issues. Bringing one of those to full expression has been chased for years by me before succeeding, but I know of several guys, and one girl, that have done it in their first year of culturing. There is no "recipe" available for finishing a healthy plant, except experience, hygiene, notes, and devotion. But once its done the work is finished. On to experiments and Frankenstein's, or shelves of cuttings in animated suspension, or even encapsulated artificial seeds, roses with thc!

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Mold in the sample vessels is what plagues most home culturists sometimes. For years it was a numbers game, take 100 explants, and some will fail, some will mold, but some thrive. each time you wish to make a change to the explant sample the risks begin again.

Then I bought this killer lcd 1600x microscope at an auction and saw the errors in my ways. The sonic toothbrush cleaner solved half of the problem by really scrubbing the samples the way they needed. But the real find was the sources of the different kinds of molds I got real good at growing. The microscope let me see the small hairs, or skin bits etc that were falling into my mini sealed greenhouses(baby food jars=bfj). these were the sources of molds introduced to the vessels, ultimately overcoming the living plant dna and colonizing exponentially. Now I wear full face arm and hand protection for the win.

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ilynnboy, on 20 Oct 2014 - 08:51 AM, said:

Wow grass,

I always wanted to do a six pack plant.

that was awesome.

Cloning

Grafting

Tissue culture

What's next?

Can you combine two sets of DNA and one petri dish and see what you get?

I am going to do some reading on tissue culture and get back to you

this excites me.

I well did the grafting go?

How many cuts did you make to get all six to take on the grafted mother?

That is something I've always wanted to do

 

 

watch the threads closely, gonna turn the haters on to the real deal pest management protocol out of state commercial growers are flocking to.mine

 

 

Yes, two sets of cannabis dna can be combined in vitro for a pollen less breeding of sorts. There is a membrane on the dna(protoplasm) that must first be breached.

 

 

 

 

 

I chose a healthy White Rhino as the host. I started with a LV Lemon cut at the base of the host plant. when that healed a month or so later I added more. Some times the first attempt would succeed, others took a dozen. always mold man, that's the enemy. At least in culture steps can be taken to sterilize plant material for use, but not for whole plants. bleach and 71% alcohol became my friends. I was able to support indica, sativa, and hybrids this way with one host.

 

 

Unfortunately I was not able to succeed in grafting auto's, an obsession of mine yrs ago. Grafting was maybe my most fun I've had with cannabis, except quality control of course, with product sampling being my forte ! lol

 

 

 

There are different approaches to clonal storage of cannabis. If not familiar with the culture culture, then a culture(had to) would mold the first time you fussed with it. But that one will last a hundred years I suspect, with some maintenance. they would need a small light source, a warmer environment, and some pest controls in the culture area. Yep, pests are huge problems, seen AFTER you have the rest of the sterile technique figured out. These little mites(yep, frikken mites) will sense the sweet sugars in your bbf jars and they will find a way in. when they do, they will poop, and it will mold, or they will bring in dust that molds too. spray glue entrances, tacky strips on the floor, shelving risers, etc go along way at prevention. these will not survive in the fridge.

 

 

 

 

 

Or a cutting could be sterilized and stored indefinitely in a test tube. It could be removed ten years later, looking the same. The end will be knotty, and need to be cut and scraped, then rooted in rockwool/cubes, or more cutting s could be taken from it, resterilized, and re stored. This will survive in the fridge for a couple weeks, but needs to be with the other cultures for best results.

 

 

 

 

 

Here's a teaser--- you want root cultures. carrot culture mix will suffice, until ready to grow.

 

 

someone could do it for you, but the sample would be useless in a few months unless you knew how to support it.

 

 

transferring to new food sources, new jars, etc and keeping them sterile can be a numbers game. I used to start with a hundred test tubes to

 

 

get a dozen still going next year. Now I can culture in a couple months, with full production(if I could) in a couple more. Maybe I was supposed to be living

 

 

in CO?

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