Marijuana Tissue Culture Success!
- Thread starter pharmacoping
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Dr. Tissue Culture
Member
You will be buying it soon...Trust me...And 1 million you say, you must be one of the shitty dutch seed kings, no thanks, I only put out the best products....
You will be buying it soon...Trust me...And 1 million you say, you must be one of the shitty dutch seed kings, no thanks, I only put out the best products....
Assumptions... cute. You seem a bit defensive. With 1 million in vitro you must make 30,000+ stabs/day and sell at least 15k. That's over 60 stabs per second during a workday. Certainly you can show a pic of your factory line of at least 150 people doing stabs, right? That's an annual revenue of about 0.2 billion $CAD. You should be out getting your pewter polished not on the forums polishing it yourself. Why does your website have nothing in it? Do you really think the market will bare those prices? Have you ever been in the starts business? Are you culling 80% of your line this week like other large starts operations? Certainly you know why, right? I could have traded part of my bank for your 120 (which is not much btw). I'm still willing to trade you 2 for 1, and only if the cultivars are decent. PM me and you will have 60 new verietals at your doorstep. I'll even take BC grade stuff. I have the cannabis cup clone only sets. Don't let ego get in your way. The TC community is a small one, and you aren't the big dog on campus yet, but I wish you luck (you will need it with that attitude)
No. You will destroy cells with liquid nitrogen. A liquid nitrogen freezer will keep the temperature to 77K which is in the right ballpark for cryopreservation. You need to use DMSO or a glycol solution, autoclavable microcentrifuge tubes (cheap, small, good for callus culture) -- drop your material in the cryopreservation medium and toss it in your nitrogen freezer. Gloves recommended. See "Plant Cryopreservation: A Practical Guide" which is everything you will ever need to know.
Dr. Tissue Culture
Member
Your numbers are slightly off my friend, and to be honest, my numbers are actually pretty conservative, as far as pics of my lab, yes you will see very nice pics in the May issue of Skunk Magazine...
...What i have you can't buy anywhere in the world, and when the magazine story comes out you'll see exactly what i mean...And as for the BigDawg in town, yes i believe i am the biggest dawg in the cannabis tissue culture world, all the big names know exactly who i am as i have been doing business privately the last few years with them...If you want to chat with me, like i said to many others, let's talk shop and you can Skype me, or come meet me in person at the treating yourself.com Metro Toronto Expo at the Convention Center on May 26th...And as far as the market baring these prices prices, people will happily pay that as they have done for the last few years purchasing from me privately here in Canada...If you think my prices are expensive, geez i wonder you must think of the dutch shit thats out there for 150 dollars..
jkahndb0
Well-Known Member
If you preserved an MJ TC in Liquid N[SUB]2[/SUB], would it last Indefinitely?
Ive heard it both ways...LoL
(III. PRESERVATION AND STORAGE. Liquid N[SUB]2[/SUB] is used to preserve tissue culture cells, either in the liquid phase (-196°C) or in the vapor phase (-156°C). Freezing can be lethal to cells due to the effects of damage by ice crystals, alterations in the concentration of electrolytes, dehydration, and changes in pH. To minimize the effects of freezing, several precautions are taken. First, a cryoprotective agent which lowers the freezing point, such as glycerol or DMSO, is added. A typical freezing medium is 90% serum, 10% DMSO. In addition, it is best to use healthy cells that are growing in log phase and to replace the medium 24 hours before freezing. Also, the cells are slowly cooled from room temperature to -80°C to allow the water to move out of the cells before it freezes. The optimal rate of cooling is 1°-3°C per minute. Some labs have fancy freezing chambers to regulate the freezing at the optimal rate by periodically pulsing in liquid nitrogen. We use a low tech device called a Mr. Frosty (C#1562 -Nalgene, available from Sigma). The Mr. Frosty is filled with 200 ml of isopropanol at room temperature and the freezing vials containing the cells are placed in the container and the container is placed in the -80°C freezer. The effect of the isopropanol is to allow the tubes to come to the temperature of the freezer slowly, at about 1°C per minute. Once the container has reached -80°C (about 4 hours or, more conveniently, overnight) the vials are removed from the Mr. Frosty and immediately placed in the liquid nitrogen storage tank. Cells are stored at liquid nitrogen temperatures because the growth of ice crystals is retarded below -130°C. To maximize recovery of the cells when thawing, the cells are warmed very quickly by placing the tube directly from the liquid nitrogen container into a 37°C water bath with moderate shaking. As soon as the last ice crystal is melted, the cells are immediately diluted into prewarmed medium.)
Ive heard it both ways...LoL
(III. PRESERVATION AND STORAGE. Liquid N[SUB]2[/SUB] is used to preserve tissue culture cells, either in the liquid phase (-196°C) or in the vapor phase (-156°C). Freezing can be lethal to cells due to the effects of damage by ice crystals, alterations in the concentration of electrolytes, dehydration, and changes in pH. To minimize the effects of freezing, several precautions are taken. First, a cryoprotective agent which lowers the freezing point, such as glycerol or DMSO, is added. A typical freezing medium is 90% serum, 10% DMSO. In addition, it is best to use healthy cells that are growing in log phase and to replace the medium 24 hours before freezing. Also, the cells are slowly cooled from room temperature to -80°C to allow the water to move out of the cells before it freezes. The optimal rate of cooling is 1°-3°C per minute. Some labs have fancy freezing chambers to regulate the freezing at the optimal rate by periodically pulsing in liquid nitrogen. We use a low tech device called a Mr. Frosty (C#1562 -Nalgene, available from Sigma). The Mr. Frosty is filled with 200 ml of isopropanol at room temperature and the freezing vials containing the cells are placed in the container and the container is placed in the -80°C freezer. The effect of the isopropanol is to allow the tubes to come to the temperature of the freezer slowly, at about 1°C per minute. Once the container has reached -80°C (about 4 hours or, more conveniently, overnight) the vials are removed from the Mr. Frosty and immediately placed in the liquid nitrogen storage tank. Cells are stored at liquid nitrogen temperatures because the growth of ice crystals is retarded below -130°C. To maximize recovery of the cells when thawing, the cells are warmed very quickly by placing the tube directly from the liquid nitrogen container into a 37°C water bath with moderate shaking. As soon as the last ice crystal is melted, the cells are immediately diluted into prewarmed medium.)
Dr. Tissue Culture
Member
Your willing to offer me 2 for 1? No thank you, why? most likely it had a Dutch strain in it, we will not deal with anything shitty dutch...
...Of course as you already know, 99% of all commercial strains out in public have at least 12% shit dutch...what does this mean? It means that GW Pharm. will sue anybody and anything who sells there shit for starters as they own the patents to all the so-called dutch shit thats out there...dont believe me? do your research and find out...by the end of the year we will have over 10,000 unique original strains that date back 35 plus years that we have patented with it's Genome...but like i said, if you like to chat further on these matters Skype me and we'll chat...and by the way, maybe you or someone else can call GW Pharm. and see if they might have heard of me and see if they might not be a tad worried...Don't get me wrong, i think your knowledge sounds better than most on this forum, but you know as well as i do that tissue cultured hardened rooted plants is far better than what is currently out there presently, it's up to me and knowledgeable people like yourself to educate everyone on the benefits of tissue culture and how it can improve everyone's life and grows...also, 1 million in vitro is not that big my man, there are plenty of tissue culture labs in the world with over 10 million per year...
Your willing to offer me 2 for 1? No thank you, why? most likely it had a Dutch strain in it, we will not deal with anything shitty dutch...
Don't get me wrong, i think your knowledge sounds better than most on this forum, but you know as well as i do that tissue cultured hardened rooted plants is far better than what is currently out there presently, it's up to me and knowledgeable people like yourself to educate everyone on the benefits of tissue culture and how it can improve everyone's life and grows...also, 1 million in vitro is not that big my man, there are plenty of tissue culture labs in the world with over 10 million per year...
I fully recognize that 1M cultures is not many, but most labs have only a couple of customers. I hope you're using a distribution chain.
On the subject of varietals and intellectual property: in the United States you may only patent an asexual plant that cannot reproduce sexually. Also, you can't patent a plant that is found in nature. It must meet the statutory requirements of novelty and all of the shenanigans referred to by the general populous regarding Monsanto and genetic marking is irrelevant. For one, Monsanto did strong arm some midwesterners but had no legal right to do so. Monsanto also won some international cases that were absurd, but these new rules were clarified and unified in 2004 by the Uruguay Round Agreement if I recall correctly. Canada and the US share IP laws by NAFTA so I am confident that there would be nothing but frivolous lawsuits should the issue of IP law arise. I have a doctorate in a life science, degrees in international business, patent law, and engineering. I would be the authority on this issue in other words.
As for the varietals that I have available.. well, you are sorely mistaken. The very talented breeders/collectors are in their mid 60's now, and they have seeds that they haven't cracked for 3 decades... that is, until us in-vitro guys came around. I have a set that was made collaboratively with Amsterdam but most of mine are not. I work with collectors/breeders so I carry breed stock, very little washout (mostly washed out cannabis cup "winners"). I'm looking for Kong. I'll trade a dozen clone only cannabis cup winners for a real, original Kong pheno. It's the only Canadian that I'm truely seeking right now, but I don't have a large Canadian collection. I'm also looking for that old strain line from circa 1980 that smells like hot garbage. The original landrace was black with bright red hairs. If you were a breeder you would probably find my breed stock by word of mouth. Guess I'll wait for you to come to me
thump easy
Well-Known Member
im gona do this culture sample do you think i can travel with it on the plane to colorado.???? my friend lives out thier and i have been wanting to vistit him also take him my jenetics. is this posible and will costoms find it on me should i take it to the hoop? lolz just kiding can i mail it out?
pharmacoping
Active Member
or, you could put it into a triple wrapped condom, swallow it, and retrieve later, but not much later..
lolz-ship it
lolz-ship it
Do most people just TC so they don't have to deal with mom plants?
thump easy
Well-Known Member
were can i buy affordable equipment?
thump easy
Well-Known Member
come on pros i dont want to read the past i need info on the now of days kinda set up.?
thump easy
Well-Known Member
you fuckers made me read!!!!!!! lazzy ass!!!!!
ajgrowsalot
Member
Is any of this even necessary? If you need so many plants you are worried about plant numbers you probably aren't growing "medically" anyway so why concern yourself with staying within medical numbers. I just seems so unnecessary to me to be growing plants in test tubes when they are so easy to grow in the first place. Sure you have to have a mother plant, then take clones (oh, how difficult) but it still seems easier, cheaper, and faster.
The science is cool but the kits that Billy Mays is talking up just seem silly.
The science is cool but the kits that Billy Mays is talking up just seem silly.
connoisseurde420
Well-Known Member
its to keep integrity in stock cloning over and over if anything happens to that mother then ur done making all those people happy..
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