Marijuana Tissue Culture Success!

Guile

Active Member
For those too lazy to read the whole thread, I DO NOT HAVE ANYTHING FOR SALE. I am not selling anything to you. i WILL NOT SELL TO YOU, OR YOU EITHER. i HAVE NEVER MADE ROSES WITH THC, OR TENDRILS. I HAVE NEVER INSERTED A BIOLUMINESCENCE GENE INTO MARIJUANA. i HAVE SEEN IT DONE THOUGH. i WAS NOT ALLOWED TO PHOTOGRAPH THAT. I AM NOT EXPERIMENTING WITH TISSUE CULTURE ANYMORE. I AM NOT TRYING NEW THINGS WITH TISSUE CULTURE. I HAVE BEEN PERFORMING THE SAME PROCEDURE, FOR THE SAME PURPOSES, FOR MANY YEARS. i AM SUCCESSFUL, AT TISSUE CULTURE,MARIJUANA CULTIVATION,MARRIAGE, FATHERHOOD, AND HAVE A REALLY BIG DICK TOO, IN CASE YOU WONDERED....i MIGHT EVEN HAVE A PICTURE OF IT SOMEWHERE...YOU KNOW, FOR PROOF AND ALL.... LETS SEE.... OH YEAH, MY AVATAR IS THE FLOWERED STRAIN OF PLUSHBERRY, AND DID SPEND HER EARLY LIFE IN A JAR, SHE ACTUALLY SHARED A JAR WITH 14 OTHER LOVELY LADIES OF THE SAME. THEY ALL GREW NEARLY IDENTICAL, AND THE HIGH IS EXQUISITE. i DID NOT CREATE HER, ONLY NOURISHED AND HOUSED HER. i DO THANK NATURE DAILY, AND SO SHOULD YOU
Hey man, I hope you're not bent out of shape.. I'll offer you a perspective, and hope it helps..

I'm a disabled vet, I got all the time in the world on my hands and even a little money coming in to amuse myself. What amuses me is to learn new shit and see people smile. (smiles are the currency of love, and we need to boost that economy)

I pick up all kinds of rediculis projects to amuse myself or make something useful to others. Recently the best parts of the world of marijuana just became available to me.. I've seen (via the internet and personal experimentation) more in the last 6 months than I had in the 15 years of independant growing before that. Unfortunately most of what I've seen seems just impracticable for me. I give away most everything I make, so I can avoid having to answer for everything I do (the feds still frown on this stuff). and my pension doesn't cover much more than my living expenses (not with my current light bill). The truth of the matter is that this has not turned out to be a profitable hobby, but I love it... because the amount of smiles it brings and the kind of people I meet..

So when I hear that one of these super interesting out of reach things is as simple as I would like it to be.. Its just easy to feel skeptical, I apologized for that, I think some others would too if they have an experienced sort walking them through it using established and well proven methods that can be reproduced in the home under reasonable measures.

I would honestly appreciate your incite and any wisdom you are willing to share, though I do tend to debate for the sake of an education (I learn alot from questions, and the responses I get from speculation).

Don't let what people think of you get you too down... They don't know you as well as you do... You can chalk it up to ignorance right? (they just don't know any better yet).
 

pharmacoping

Active Member
Guile,

nah, not bent at all, and appreciate your candor, wit even. so we start out with the kids not believing, then believing, but not admitting the possibility...I just cant convince someone they can walk, then teach them to run, just so I can go jogging. a set of instructions is in the links my friend, if there are any q's about them, ask me, maybe I can help. I sense the only genuine interest so far.
heres some more great links'
Recently, the genetics team at DinaFem Seeds has released an autoflowering strain that they have spent some time perfecting. It is said that,

"For anyone who is a fan of sativa-dominant hybrids and hasn’t tried growing an auto-flower strain, DEFINITELY pick some of these seeds up. You won’t regret it!"

Road Runner was developed after much genetic engineering and trial and error. Throughout its life, its growth is astounding for being an autoflowering strain and is said to have a very easy trim at the end of its 90 – 100 day life cycle. Its friendliness and harvest rewards are greatly hyped up making this an excellent strain to experiment with.

keep in mind, genetic engineering takes place when you pollinate by hand. most pants have internal virus' all the time, the knots in th estalk are groupings of these virus, and the protective cells keeping it at bay, they have a name, but its a secret(lol)
buy Plants from Test Tubes for 20 dollars on amazon and you'll arrive at tissue culture 101 in a week. pressure cooker, hormones, baby food jars, lids, agar, food mixes. a magnetic stirrer is a blessing, heated would rock, but not necessary. nother secret, dmgold Replicator will perform a small miracle in a sterile test tube with agar and sugar, nutes, for beginners, 1ml of Replicator to 1 liter of mix. I dont use it anymore, as I have pure hormones/amino acids to work with now. gelatin gave me my eye opening experiences a long time ago, might have some luck on the cheap.​
 

Gastanker

Well-Known Member
I guess I should also apologize. I don't like new comers to the site trying to sell products based on absolutely ridiculous claims. I understand that you are no longer going to try and sell your kit and that you have retracted some of your ridiculous claims so by all means show us how you do it. I would be legitimately interested in a step by step preferably with pictures to make it easier to follow along.

Pharma - how long does it take you to go from tissue culture to a 3" tall normal looking plant? From what I have seen it takes 4+ months for a cultured marijuana plant to reach the size that it can leave the beaker and another month or two to get it to the same stage as a standard clone. For most of these months the culture still counts towards plant numbers - it counts as a plant the second it has a recognizable leaf/root system regardless of whether its hydro, soil, aero, or in a petri dish. Like you say a great way for long term stasis or if you want to create thousands of identical plants at a time (you would of course need to be able to legally grow thousands at a time), but for the typical producer - even really large producer, it's not practical.


@ Guile -

Great video how to on plant tissue culture with step by step procedures. http://www.youtube.com/user/fbt2007

Kits are cheap ranging between $30-$250 depending on if it's simply the hormones or containers and tools as well. Tissue cultures are a blast and I would recommend anyone seriously into horticulture to at least experiment around with it. I would recommend starting with African violets as they are extremely easy. Orchids and pineapples like in the videos are fun as well and not to complicated but marijuana is much more difficult and really not very practical.
 

pharmacoping

Active Member
sure thing man, but I made no ridiculous claims, except that I would assemble and sell kits, thats ridiculous, so maybe we should ge back to that also, because it will be a chip on your shoulder throughout your experience with me, and I did not lie. If you've read every link I've posted, you would have seen this tech in play. I keep callus of different species in suspended animation. It takes a few weeks to get a good chunk of callus going. When it looks the size of a lima bean or so, thats when I cut it into four, transplant to divide each. These callus replicate within 3 days. I cut them into four again, if plants are needed, mind you. If not I just let it naturally replicate until needed. still no sign of plants. the needed ones will be transplanted into a rooting hormone/sugar/gel/nute mix and begin rooting in a day, a test tube will fill the bottom up with roots solid in three, still no plants. heres the magic, after those rootballs(which can be divided also with great success)are transplanted into a cytokin/auxin mix, causing wicked like lateral branching and upward bonzai growth in a couple days. answer in short is I tend mini plants in containers for app. 2 weeks(two inches, nowhere normal looking until a couple weeks into veg and the leaves spread out and all shoots up, with some luck. they harden in veg, and continue there for exactly 45 days, to flower, which is 60 days for most of my herd. how can you instruct an entire forum that mj is complicated and not very practical? thats a bold claim dude. roses with thc , and way more interesting stuff goes on while you get high. There are plants growing human tissue man, and you balk bioluminescence, or, play some catch up, read a bit more, and when you realize that bioluminescense,carniculture,roses with thc, or even tobacco, or even hundreds of plants with thc,are not wild claims, and are actually part of history. then, come back and request I babysit a step by step with photos, and an audience who currently thinks this is impossible and cant be done, get real man. wrong thread for you, really.try this, maybe you'l understand http://academicsreview.org/reviewed-content/genetic-roulette/section-2/2-2-plant-tissue-cultures/good luck, no pics, or directions till you read suggested materials, and get our equip !you saw experimental failures with many untold variables. they molded, we know that, they all were deforme, and took months to grow. this sounds like me, 15 yrs ago with jello,vitamins,and epsomsalts. yep you're right, it cant be done.geesh
 

jeffdamann

Active Member
pharmacoping, Ive been following this thread since you started it, because Ive been studying tissue culture.

It occurs to me that you should post pictures of your methods and a Tutorial on how to accomplish the same thing.
I mean anyone can come in here and say the things youve said and not offer proof, but prove the naysayers wrong and post a faq with pics, You will be repped and liked like crazy for it too, so why not?

If you cant get your hands on a camera you CAN get your hands on a cell phone.
 

Voidling

Well-Known Member
I have a paper from Chinese researchers on tissue culture of the hemp plant. I skimmed in but was a bit over my head without more in-depth study of it. Hard to read and study charts on my cell phone screen.

It is something I plan to try after getting my grow setup proper. I would love it if you would post your mixes of hormones that you have got to work for this. I'd love to scrap mothers
 

pharmacoping

Active Member
hi jeff,

i do have a camera, two actually. one i've used to post pics here already. I will post some , but I do not have photo shoot already done, so I'll shoot as I go maybe. I agree pics or gtfo, but really neg attitudes from the start, and dont have time to argue the technology with anyone. I want more, who are already on board, culturing successfully, to chime in. the links provided show films, pics, step by step, supplier lists, tc on a budget,equipment lists, past and future reports, current patents being established, and even home labs around the world, even public ones! reposting the how to's and the progressive photos would be a little redundant, and time consuming. the pics I'll post will not be from beginning to end though, as I just took my first pics. i'm not a pic posta, dont write papers, and only just now discovered my first "combative" forum. mj forums, specifically exist for mj tissue culture already...beginning to think google is only accessible by me.
on the cheap is prolly the sought after info here, so pics, equip etc mentioned by me are for the basement dwellers i guess. follow links,get good equip,do it right if you can, otherwise you get failures like mold, mutants,and 4 month old tissue cultures growing slow.

sure would like to see proof of the moldy experiments, just to mention.
 

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pharmacoping

Active Member
the "transfer lab" can be as simple as this tupperware container. this is only where cultures are divided, and transferred. a seperate area is where they grow out.Four most commonly used plant growth regulators (BAP, KINETIN, IBA,and NAA) i use ppm, a preservative....descriptions for their intent is on google. i pre mix mine to 1 mg/ml concentration, or buy them that way if it's cheaper for you.they are incredibly expensive pure. keep in mind a couple drops of that 1mg/ml will show results in a 1 liter mix of sugar,gel,nutes,water. less is better than more, always. scalpels, tweezers, hand held microscope maybe. paper towels. sterilize it all in a pressure cooker for 25 minutes highest pressure. always let it cool for many hours before opening. sterile protocols apply hereafter. many auxins, cytokins acids, bacteria, etc are available to express cells in various ways. every single one is explained in great detail in the links provided.

I am getting familiar with suspension tissue culture now. this is a procedure that involves suspending a tissue above the mix, rather than in it, draining,adding a new mix, all without a transfer. saving bucks and time. much more involved, but super cool.
 

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pharmacoping

Active Member
hmm, noticed that some here are already active with every hormone used, even cloning with them. much could be learned from their experiences.
 

Gastanker

Well-Known Member
roses with thc , and way more interesting stuff goes on while you get high. There are plants growing human tissue man, and you balk bioluminescence, or, play some catch up, read a bit more, and when you realize that bioluminescense,carniculture,roses with thc, or even tobacco, or even hundreds of plants with thc,are not wild claims, and are actually part of history. then, come back and request I babysit a step by step with photos, and an audience who currently thinks this is impossible and cant be done, get real man.
O shit - you are STILL claiming that labs across the world (most of which cannot legally experiment with cannabis) are wasting their time making THC laden roses and glow in the dark pot plants? I'm curious as to why a high tech lab would ever attempt such ludicrous experiments. And that you have personally seen them... :shock:

Are you saying you are unwilling to share with this community how you grow out tissue cultures? I'm really really confused. Why did you post here initially if not to teach this community what you have learned.

This is equivalent to the proof you have posted. Look tissue cultures!



Look pot plants!



I'll be honest - none of these pictures are mine and are completely unrelated. You have yet to post a single picture of a discernible cannabis tissue culture - you realize that right?
 

pharmacoping

Active Member
gastanker

I suspect that most of the links and information I've shared is discernible to you, also. If you just would read "Plant From Test Tubes" your head may miraculously fall right out of your ass, as long as the text is discernible, to you. I have not seen roses with thc, I have seen a mj plant with the bioluminescense gene inserted in vitro creating a mutant strain of dna within the emerging plant. It was grown, and did flouresce(sic?) the labs around the world are not focusing on roses with thc, but are being granted government patents monthly for many mutagenic marijuana creation. They may alredy have that one too, I dont know.

Do you accept the well known fact that plants are currently growing human replacement tissues. corn,wheat,rye,soybean etc have been grown with a mutagenic strands that are responsible for things like "round up ready" or corn with pesticides inside there genetic makeup.

I've been asked for photos, and have taken some. I am not a pic taker, nor have I photos of my gardens. I am giving photos as I take them, and now see these too, are unacceptable, as suspected, and predicted.from your comments alone, we can tell that you have not bought a book on the subject yet. Please do, as you can join the enlightened at that time.


that is a big tc operation man ! nice pic. I stay within a closet, a few dozen vessels, and one bank of t5 lights for culturing. never manipulated in strange ways, and use the system purely to avoid plant count issues. cloning is easier, but takes up a number as soon as its in rockwool. stored growing roots are a huge head start and when told to vertically shoot imagine the advantage to a ball of roots the size of a golf ball at day one. i'm not a mad scientist, or trying for patents, but HortiPharm is, and three others backed by Bayer, Monsanto, and some big pharma corps. check into it, you might find roses with thc by now? Hops is a shoe in for this procedure, as it's the same genus, or something,dude, uha,uha
 

Gastanker

Well-Known Member
I never once denied genetic engineering nor tissue cultures... I do refute your claim of THC laden roses and that you have seen a bio-luminescent cannabis plant. I've worked in several labs and I have an extremely hard time picturing a scientist fucking around for the sake of fucking around with a plant that can get them into loads of trouble. Three times now you have stated that there are in fact (as if you have personally seen them/know of them) roses with THC. Now you are saying there aren't but there "could be"? The president "could be" an alien...

I have stated that in my opinion tissue cell cultures do not make any sense to most marijuana farmers due to expense, time, and there are very few true benefits. From your own link here is the time scale for tissue culturing cannabis plants -



According to this there is a 2-6 month period before the culture can be transferred to soil at which point you still have a very long veg time due to the minute size of the plant. I assume the author was a Phd? With his high tech lab with full autoclave... he still had contamination issues.

Again I'm curious as to your process - can you explain how you personally do this? I understand others can tissue culture pot, I understand that large corporations are genetically modifying a huge assortment of different organisms... You said you had mastered the tissue culture - please share with us your procedure.
 

pharmacoping

Active Member
after wondering why I haven't met anyone here with mj culture success, a possible answer dawned on me. This is the most combative,immature, misinformed marijuana community I have ever encountered. Some of these same mindsets are, fuckit, ..

I thank the couple that helped me find two forums full of like minded individuals successful in the art(evidently) of marijuana tissue culturing, botany, and horticulture. I am a member in these communities now and will probably never return here, well, unless I'm having a hard time figuring out if my female has been seeded(brown pistils two weeks flower, as suggested here by a member) or if its a male or female.
 

pharmacoping

Active Member
SEEDS ? i never thought of thaT

chart is spot on.
heres the missing part though
once all these jars are set up, none are plants yet mind you, then each jar of "massive root growth" can be "planted" and sprouts within two days, grows fast from theer, virus free, and faster than any clone/seed, as it already has a ball of roots. they count as plants only in the last couple days of sitting on the shelf, dividing, making more rootballs until ready. one seeds' roots will turn into hundreds of seperate rootballs of identical genes in a month. each super start is ready when you are. so you see its not the time to extablish that is important, as they too are now part of the perpetual experience, being transplanted weekly.

marijuana has been dna sequenced(last month high times) and the patent gold rush is on(thats why labs can play now)

seems failures fare better than successes here. a pic of my first tissue culture freak, and a pic of my double chromosome PlushBerry, colchicine success. sorry it looks so good, I tried to take a shitty pic, oh well
 

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ddimebag

Active Member
I read up a bit on genetic modification using agrobacteria...to successfully create a transgenic rose (or any other plant except cannabis) that produces THC, one would first have to figure out exactly what biochemical processes are responsible for the formation of THC, determine the genes that control the production of the necessary enzymes, and control the mechanism by which these enzymes interact with their substrates (also what those substrates are and where they come from), obtain or create the necessary restriction enzymes to cut the DNA at precisely the right places, introduce the cut up genes into a DNA plasmid of the agrobacteria (a process for which one would need more custom-made enzymes), allow the bacteria to infect a tissue culture of your transgenic plant, and then hope that the new genes will get integrated into the plant DNA. All of this has to take place in sterile laboratory conditions, and even then, there is a significant chance of failure ( for example, from infection). I seriously doubt that any lab has actually managed to perform such a feat...or even tried to...I have seen and read about transgenic glow in the dark tobacco plants (which was done as a proof of concept, and only required the addition of a single gene), and I have performed this type of genetic modification on E. coli bacteria, also with a single gene. While this type of genetic modification is possible, making roses with THC is, for now, science fiction (as far as i know).
 

ddimebag

Active Member
using tissue culture to get around plant number restrictions is interesting though...i would like to read more about that...
 

Voidling

Well-Known Member
I read up a bit on genetic modification using agrobacteria...to successfully create a transgenic rose (or any other plant except cannabis) that produces THC, one would first have to figure out exactly what biochemical processes are responsible for the formation of THC, determine the genes that control the production of the necessary enzymes, and control the mechanism by which these enzymes interact with their substrates (also what those substrates are and where they come from), obtain or create the necessary restriction enzymes to cut the DNA at precisely the right places, introduce the cut up genes into a DNA plasmid of the agrobacteria (a process for which one would need more custom-made enzymes), allow the bacteria to infect a tissue culture of your transgenic plant, and then hope that the new genes will get integrated into the plant DNA. All of this has to take place in sterile laboratory conditions, and even then, there is a significant chance of failure ( for example, from infection). I seriously doubt that any lab has actually managed to perform such a feat...or even tried to...I have seen and read about transgenic glow in the dark tobacco plants (which was done as a proof of concept, and only required the addition of a single gene), and I have performed this type of genetic modification on E. coli bacteria, also with a single gene. While this type of genetic modification is possible, making roses with THC is, for now, science fiction (as far as i know).
It sounds like it would be technically possible just unlikely anyone has done it.

On the one hand, genetically modifying things seems really neat and cool, something I might would even go to college for. On the other hand, I'm kind of against the gmo's and monsanto. I try to use the organic seeds in my garden for veggies.
 

ddimebag

Active Member
It sounds like it would be technically possible just unlikely anyone has done it.

On the one hand, genetically modifying things seems really neat and cool, something I might would even go to college for. On the other hand, I'm kind of against the gmo's and monsanto. I try to use the organic seeds in my garden for veggies.
Yes, it is TECHNICALLY possible (and VERY expensive...each of those restriction enzymes costs hundreds of dollars per ml), but because of legal, patent and profit issues, i don't think anyone has done it yet, or will be doing it any time soon. Also, imo, people who are against genetic modification are simply scared of something they do not understand. If you have an interest in this area, you should definitely study it in college; it will give you a much clearer outlook on life, what it is and how it works. Biotechnology is here to stay, so there will always be jobs in that sector...
 
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