Colorado Sam
New Member
Being a newbie, my only question is, how does the plant function leaves? Not accusing, curious.
Marijuana Tissue Culture Success!
- Thread starter pharmacoping
- Start date
Being a newbie, my only question is, how does the plant function leaves? Not accusing, curious.
chemtrailsrbad
Well-Known Member
Fuck you and your genetically modified shit!
im not sure what you mean exactly but a plant in tissue culture doesnt do a lot of photosynthesizing. you provide the culture with sugar to provide most of its energy.
your a dumb motherfucker that has no idea what hes talking about
pharmacoping
Active Member
To the believer, no proof is necessary. To the nonbeliever, no proof is sufficient.
pharmacoping
Active Member
Culturing is not much different than your grandmother sticking a bit of broken stem from a favorite spider plant into a glass of water on the windowsill. we're storing plant material for long periods of time and some sterile technique measures are needed to ensure viable mold free samples to work with. You can experiment in a similar way with cloning, gel, and a block of rockwool. The plant will ultimately develop roots, and begin its life once again. Our favorite plants are asexual and able to reproduce the whole using only a specimen . YOu could experiment with a healthy leaf snip half tucked into a sterile rockwool cube, moist with a bit of cloning gel, and you may see roots or even chutes in no time at all. Its a very simple, advanced method of propagation. Tissue culture will not replace simple cheap methods of cuttings and clones, but yet is exactly how 99% of our vegetables are stored, produced, modified, or otherwise propagated. Bananas, venus fly traps, and many flower specie are delivered to the nursery as cultured samples ready to plant chute and root, with some love of course.
pharmacoping
Active Member
[h=1]Living, breathing, glowing rabbits successfully born[/h]
Two out of eight bunny siblings got the glowing gene.
It sounds more like a child's toy, but glow-in-the-dark bunnies are now a living, breathing reality -- and researchers say they could help bring affordable drug therapies to developing countries.
In a collaborative effort between the University of Hawaii at Manoa and two universities in Turkey, scientists created two transgenic (genetically modified) bunnies by inserting a gene from jellyfish DNA into a rabbit embryo.
Furry and white in normal lighting, the pair glows bright green under black light. These are the first transgenic rabbits; transgenic mice, cows, goats, chickens and pigs have been successfully produced.
"Because I've been around it so long, it's almost become second nature," UH Manoa's Stefan Moisyadi told CBSNews.com. "But the rabbits were sensational, they were glowing so bright that even though their fur [was thick], it was even shining right through the fur."
The rabbits were conceived via in vitro fertilization (IVF). The researchers first extracted embryos from an adult female rabbit. Next, they inserted plasmids into the embryos, in a process similar to that developed for human IVF by Moisyado's mentor, Dr. Ryuzo Yanagimachi.
Founder of the school's renowned Institute for Biogenesis Research, Yanagimachi famously cloned adult mice in 1998, and was an early pioneer in the development of IVF. Originally, IVF was passive, as scientists had to wait for the sperm to swim to the egg. Yanagimachi further revolutionized it by developing an active process in which the sperm is inserted directly into the egg. The method is now used in fertility clinics around the world.
To transfer genes of one species into the embryos of another, Moisyadi essentially swaps the sperm from Yanagimachi's method with plasmids. The plasmids contain enzymes, or transgenes, from the jellyfish DNA as well as a tool, called a piggyBAC, that transfers the transgenes. The piggyBAC is designed to disintegrate once it's completed its role. The embryos are then injected back into the mother. The entire process takes less than half an hour, but there's no way of knowing the outcome until the mother has gone through a regular pregnancy.
The fluorescent protein from the jellyfish DNA has no use aside from telling researchers if their experiment was a success. Basically, they wait until the offspring are born, then flick off the lights. If the pups glow under black light, it signals success. "It's just a marker, it has no value whatsoever," explains Mosiyado.
While cool to look at, there's more to these transgenic animals than meets the eye.
Now that they have proven the success of active transgenesis in rabbits, the researchers will start to use the method to inject other genes. These will be genes that produce proteins that are used in common drugs. The proteins will be present in their milk and extracted from there. The host animals don't benefit, or suffer, from the introduction of the genes. The beneficiaries are people who need access to low-cost drugs.
"You can express the protein that's made from the gene and generate large quantities of it easily and cheaply. Think of a factory built under sterile conditions, it costs a heck of a lot of money. Try doing it in Uganda, countries like that. It is less expensive to purify something from the milk."
The animals can, in essence, replace the costly labs and the cell culture methods through which proteins are currently engineered. They can function as "biofactories."
"Once you create the animals that have a gene in them that they can't make, a bioprotein, you can breed these animals and perpetuate it," Moisyadi explains.
In 2009, the Food and Drug Administration (FDA) approved the drug ATryn, created using protein from a transgenic goat, for patients with hereditary antithrombin deficiency (HD) -- a rare blood clotting disease that afflicts about 1 in 5,000 Americans. The European Union had approved the drug in 2006. GTC Biotherapeutics in Charlton, Mass. developed ATryn using protein derived from a herd of 30 transgenic goats.
Scientists at AgResearch in New Zealand are using proteins derived from the milk of transgenic cows to develop drug therapies for multiple sclerosis. The cows were not fluorescent and did not glow in the dark -- that was only used to test Moisyado's method. Rather, they were created using transfection, or the introduction of DNA into a cell. The process takes about seven days in culture.
Milk or meat products from transgenic animals are prohibited from entering the animal or human food supply. They are intended strictly for scientific purposes.
Moisyadi continues to work on the technical aspects of the active transgenesis procedure, hoping to improve the success rate to at least 50 percent. In the rabbit experiment, the method was 25 percent successful, with the gene catching on in two of the litter's eight bunnies. Other methods pale in comparison, with rates of less than 5 percent.
This rabbit experiment was actually a sideline project, while the researchers wait on the birth of the world's first transgenic sheep. Earlier this month, they injected 68 sheep embryos, and 10 have taken. But sheep gesticulate for about four months. Rabbits only take about a month.
They are expecting the first of the lambs in November, but won't know if the process was a glowing success until they flick on the black light.
Two out of eight bunny siblings got the glowing gene.
It sounds more like a child's toy, but glow-in-the-dark bunnies are now a living, breathing reality -- and researchers say they could help bring affordable drug therapies to developing countries.
In a collaborative effort between the University of Hawaii at Manoa and two universities in Turkey, scientists created two transgenic (genetically modified) bunnies by inserting a gene from jellyfish DNA into a rabbit embryo.
Furry and white in normal lighting, the pair glows bright green under black light. These are the first transgenic rabbits; transgenic mice, cows, goats, chickens and pigs have been successfully produced.
"Because I've been around it so long, it's almost become second nature," UH Manoa's Stefan Moisyadi told CBSNews.com. "But the rabbits were sensational, they were glowing so bright that even though their fur [was thick], it was even shining right through the fur."
The rabbits were conceived via in vitro fertilization (IVF). The researchers first extracted embryos from an adult female rabbit. Next, they inserted plasmids into the embryos, in a process similar to that developed for human IVF by Moisyado's mentor, Dr. Ryuzo Yanagimachi.
Founder of the school's renowned Institute for Biogenesis Research, Yanagimachi famously cloned adult mice in 1998, and was an early pioneer in the development of IVF. Originally, IVF was passive, as scientists had to wait for the sperm to swim to the egg. Yanagimachi further revolutionized it by developing an active process in which the sperm is inserted directly into the egg. The method is now used in fertility clinics around the world.
To transfer genes of one species into the embryos of another, Moisyadi essentially swaps the sperm from Yanagimachi's method with plasmids. The plasmids contain enzymes, or transgenes, from the jellyfish DNA as well as a tool, called a piggyBAC, that transfers the transgenes. The piggyBAC is designed to disintegrate once it's completed its role. The embryos are then injected back into the mother. The entire process takes less than half an hour, but there's no way of knowing the outcome until the mother has gone through a regular pregnancy.
The fluorescent protein from the jellyfish DNA has no use aside from telling researchers if their experiment was a success. Basically, they wait until the offspring are born, then flick off the lights. If the pups glow under black light, it signals success. "It's just a marker, it has no value whatsoever," explains Mosiyado.
While cool to look at, there's more to these transgenic animals than meets the eye.
Now that they have proven the success of active transgenesis in rabbits, the researchers will start to use the method to inject other genes. These will be genes that produce proteins that are used in common drugs. The proteins will be present in their milk and extracted from there. The host animals don't benefit, or suffer, from the introduction of the genes. The beneficiaries are people who need access to low-cost drugs.
"You can express the protein that's made from the gene and generate large quantities of it easily and cheaply. Think of a factory built under sterile conditions, it costs a heck of a lot of money. Try doing it in Uganda, countries like that. It is less expensive to purify something from the milk."
The animals can, in essence, replace the costly labs and the cell culture methods through which proteins are currently engineered. They can function as "biofactories."
"Once you create the animals that have a gene in them that they can't make, a bioprotein, you can breed these animals and perpetuate it," Moisyadi explains.
In 2009, the Food and Drug Administration (FDA) approved the drug ATryn, created using protein from a transgenic goat, for patients with hereditary antithrombin deficiency (HD) -- a rare blood clotting disease that afflicts about 1 in 5,000 Americans. The European Union had approved the drug in 2006. GTC Biotherapeutics in Charlton, Mass. developed ATryn using protein derived from a herd of 30 transgenic goats.
Scientists at AgResearch in New Zealand are using proteins derived from the milk of transgenic cows to develop drug therapies for multiple sclerosis. The cows were not fluorescent and did not glow in the dark -- that was only used to test Moisyado's method. Rather, they were created using transfection, or the introduction of DNA into a cell. The process takes about seven days in culture.
Milk or meat products from transgenic animals are prohibited from entering the animal or human food supply. They are intended strictly for scientific purposes.
Moisyadi continues to work on the technical aspects of the active transgenesis procedure, hoping to improve the success rate to at least 50 percent. In the rabbit experiment, the method was 25 percent successful, with the gene catching on in two of the litter's eight bunnies. Other methods pale in comparison, with rates of less than 5 percent.
This rabbit experiment was actually a sideline project, while the researchers wait on the birth of the world's first transgenic sheep. Earlier this month, they injected 68 sheep embryos, and 10 have taken. But sheep gesticulate for about four months. Rabbits only take about a month.
They are expecting the first of the lambs in November, but won't know if the process was a glowing success until they flick on the black light.
grassmatch
New Member
I'd like to discuss the net effects of tissue culturing.
1) First What is the cost of your current setup? It sounds like it would be <$500 bucks, correct me if I'm wrong.
2) Do you believe that properly performed tissue culturing is MORE or LESS successful than cloning techniques? What concerns me most about TC is that I will take up a plant space with something that WON'T work out - is that a valid concern provided that the genetics I started with are successful?
3) I'd like to understand the true benefit of TS to the legal caregiver.
You say that you can go from culture to plant in three days?
1) First What is the cost of your current setup? It sounds like it would be <$500 bucks, correct me if I'm wrong.
2) Do you believe that properly performed tissue culturing is MORE or LESS successful than cloning techniques? What concerns me most about TC is that I will take up a plant space with something that WON'T work out - is that a valid concern provided that the genetics I started with are successful?
3) I'd like to understand the true benefit of TS to the legal caregiver.
You say that you can go from culture to plant in three days?
Hydrotech364
Well-Known Member
I was at a friends house a long time ago like 1980 or something and He had a USArmy study in his possession with the secret red binder and all and it detailed a Hop's plant that produced THC by Grafting.It stated that they are somehow cousin's genetically.Looked official to me.Nothing about any other genus was mentioned.
grassmatch
New Member
Seems like you could graft some MJ onto a hop plant, it'd probably work. But whhhhy =). Also even though this is all some serious weirdness, I think grafting is some different weirdness all together. Grafting being something that someone who is more interested in keeping a mother might be interested in. Graft different strains together, not a bad plan, maybe easier than TC, but you don't get all the beni's either - from what I'm reading.
Hydrotech364
Well-Known Member
Oh I don't know, Hmmmm maybe because it is a very hearty 32 ft long plant that doesn't look like cannabis.
pharmacoping
Active Member
Hiya ! to date, not counting the decades of trial and error, I could rebuild my tc closet for about 500$, you are correct. I however have a small fortune in there, collected over decades of trial and error. a trash picking culturists dream oneday? Cloning traditionally is by far the only practical choice for a caregiver today, at least in MI! PLANT COUNT 101. when a cutting is jammed into a cube of rockwool, it is now a plant. I have read that a bag of cuttings in the fridge is also counted, as its intent was to plant/sell. A seed is not a plant. a ball of roots are not plants. undifferentiated dna is not counted as a plant. I do not support hundreds of jars of tiny plants, but maybe 6 or so at a time, and those are out of the jar and acclimated quickly, sometimes within three days of their roots showing. Each is considered a plant, with sprouts or roots. Benefits of tissue culture would be subjective I think. for thought though, one chunk of dna the size of a pencil eraser nub can produce 1200 "clonal copies" per month, from one jar, if one could use that many? Cloning traditionally carries a genetic drift eventually that is difficult to avoid when cloning from a clone, and so on. Mothers are difficult to support, require light, water etc, and will most likely be subjected to pathogens and disease and pests while indoor growing long term . Hydro growers will tell stinky root ball stories of long term plants hanging in hydro. most I know would use dirt for moms, which eventually succumb. Pathogenic hijacking is one reason one clone is vastly different than the next. Not sure of others' programs, but after spending small fortunes on large breeding projects I cant risk an offspring failure. When someone purchases a venus flytrap, or banana tissue culture, about the only way one can, it will be identical to the donor plant, while a hand cutting cannot guarantee this purity. Inside each vessel many choices are available to us such as lateral growth rate, internode spacing, and more I suspect. when I get my gamma ray device I'll be able to tell you more. TC can produce tiny bonzai mj plants under three inches, but having a hundred branches, and a predetermined node spacing.
I am sorry it takes me so long to respond. I maintain a full registry full time, and man I could share some real nightmare stories....maybe I'll start a thread about Caregiver Patient Nightmare Stories hehe, some I've heard/had are really scary..
pharmacoping
Active Member
can graft mj to hops, successfully, however done in culture or on the branch, it remains a traditional graft, not capable of regeneration, or changing dna. Other plants may be able to be gmo'd and produce an array of compounds, including thc. I don't have my gamma rays yet, so that's out for me. the big guys are doing it in wild ways. There are plant seeds the army drops on mine fields, they sprout in a day, and one nighttime fly over and every mine is located, by the gmo sprouts forced to "glow in the dark" when amongst bomb constituents. I have seen some of these freaks, including gold infused glowing plants, including cannabis, with my own eyes, more than once. The jelly fish protein is readily available, and ecoli is easily purchased. the only thing after that is agrobacteria, illegal to posess in my state. A keen ey will be able to locat this bac and culture it by itself, from any forest tree infected with Crown Gall Disease, like this plant shown..There are strict patent issues involved with that technology, and although some can be performed in a culture club closet setting, I would never do such a thing, for record..
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Hydrotech364
Well-Known Member
Saw FDD do this to a plant with cloning gel. Tony Fuckin Stark can build this shit in a Cave...Jus sayin, I wont be playing with this science til it's everywhere.
grassmatch
New Member
Haha, $500 bucks, that's all it cost to rebuild eh? Sounds like my room, hey it'd be cheap, if it weren't for all the trial an error. I got away with all that by taking from and returning items to the used market - probably harder to do with T.S. supplies. Not so common.
I've heard the argument made about genetic drift and traditional cloning. I have no doubt that it's true, to some extent. More than a few people I know, including very likely myself, are hundreds/thousands generation's in with their clones. Surely it won't last forever, but the going is good at the moment.
Careful with the gamma rays
. Perhaps some potassium-40 
You looked me up eh? Well if you want to share some war stories I'm up for it
. I think most everybody in the MMMP program has got some of their own...
pharmacoping
Active Member
I'm on hundred/thousands of generations of clones/from clones, with no issue at all also. But I also am familiar with clean habits, sterile technique, and working in a sweet setting. Most grow rooms are not dialed in, and a spore of mold is all I takes to experience a never ending drift. I bet your room is pretty clean too, hence no probs in the cloning area. I just used that as one more "advantage" to traditional cloning. I would never suggest culturing to a marijuana grower, but maybe to the mad scientist/ pro hydro/ full time cannabis industrial types. It is afterall, THE way that ALL the big corps grow, and most of our edible food begins this way today.
MsCochise
Member
Pharmacoping.....I tried pm'ing you but it did not happen on my end. I do have questions for you to start tissue culture as well as purchase a kit. I've been reading up on the entire process, but if you already have a video or other info that I can donate to you to bring me up to speed quicker, is good for me. Waiting on your advise.......
Ms C
Ms C
pharmacoping
Active Member
Hi Ms C. !! I did get your pm's. Plants From Tissue Culture is the name of the culture bible. I cant sell you anything but I'll help all I can.Putting your aquarium culture start up inside of a greenhouse will present its own set of issues. consider a still, unused closet dedicated to the craft. NO traffic, no air, no plants growing, no fungus, no insects etc is key to success. I don't use hoods, hepas, or any other air filtration while culturing. I have a heap running full time in the room that the culture closet is in. That heap stays off for a day before I transfer. Your aquarium should be used for transfers, not for the jars to sit and grow also. put them below or above your area on a shelf. I use a 1x2 t5 fixture, 100watts total to support dozens of stacked culture vessels at a time.Pharmacoping.....I tried pm'ing you but it did not happen on my end. I do have questions for you to start tissue culture as well as purchase a kit. I've been reading up on the entire process, but if you already have a video or other info that I can donate to you to bring me up to speed quicker, is good for me. Waiting on your advise.......
Ms C
peace
MsCochise
Member
Pharmacoping........I just researched and found out the best method to control growth is your suggested method.... growth room / chamber/closet, however, the next best method the research states is a greenhouse. Coincidentally we are pouring a concrete slab next week to put a greenhouse on. I figured we can move the fish aquarium to the greenhouse as well to house the transfers and to be used as our "clean box". What are your views on the greenhouse for TC? Yes, we will have heat and lights in the greenhouse.
Thanks
Ms C
Thanks
Ms C