Yeah thats purdy ..purdy crazy lol peace out headbad 707
-
Happy Birthday ICMag! Been 20 years since Gypsy Nirvana created the forum! We are celebrating with a 4/20 Giveaway and by launching a new Patreon tier called "420club". You can read more here.
-
Important notice: ICMag's T.O.U. has been updated. Please review it here. For your convenience, it is also available in the main forum menu, under 'Quick Links"!
Cannabis tissue culture
- Thread starter Gray Wolf
- Start date
LittleAmsterdam
Member
I'm starting a small project with some extra cuts I have. I've purchased a small set of supplies and will post my results soon. This excites me for many reasons ! 
thefalsediviner
Member
Strains in vitro at the moment are as follows:
Cheese Quake
White Widow
Nevell's haze
White widow x Big bud
Super Lemon Haze
Super Silver Haze
Ak-48
Hippie Crippler
Sharks Breath
Great White
I use canning jars, a home made laminar flow hood, a pressure cooker, and a microwave. I'm nor ready to post any medium formulas but I will when I get this down a bit more. Which could be a few months or more.
tfd
Cheese Quake
White Widow
Nevell's haze
White widow x Big bud
Super Lemon Haze
Super Silver Haze
Ak-48
Hippie Crippler
Sharks Breath
Great White
I use canning jars, a home made laminar flow hood, a pressure cooker, and a microwave. I'm nor ready to post any medium formulas but I will when I get this down a bit more. Which could be a few months or more.
tfd
Where'd you pull your protocols from tfd?
before http://imageshack.us/photo/my-images/834/img3521u.jpg/
after http://imageshack.us/photo/my-images/511/img2185l.jpg/ it takes some time though and you have to be very very sterile;x
after http://imageshack.us/photo/my-images/511/img2185l.jpg/
it takes some time though and you have to be very very sterile;xFaith-Hope-Love
Member
thefalsediviner
Member
That's the problem, so far I've been knuckling through this, adapting from protocols for similar plant types. Once you get the sterilization down and learn how to isolate plant pathogens, it's about hormone ratios.
tfd
I'm aware...I considered specific hormone ratios as a part of a protocol. My friends pulled their protocol from Mississippi. From there it's an infinite tweak of recipes AFAIK.
LittleAmsterdam
Member
This looks promising, but how do you transplant from Stage III to Stage IV?
tag
thefalsediviner
Member
before http://imageshack.us/photo/my-images/834/img3521u.jpg/
after http://imageshack.us/photo/my-images/511/img2185l.jpg/
Thanks for that, given the look of your medium I got a good idea what ingredient I need a touch more of. May I ask how much you drop the sprout generating component when you add the root generating component? For instance, are you reducing it by half or all together?
tfd
thefalsediviner
Member
No, I have an idea what he's using to get shoot multiplication and it's a creamy white substance called coconut milk. Why are people so quick to judge? Seems kinda narrow minded to me.
tfd
tfd
Leeroy&Co:
Active member
wow 
Hey all I thought I'd chime in....just finished our micro propagation section at school, did ky14 tobacco and begonia leaf cuttings. All work was done in flow hoods, utensils flame sterilized, hands sprayed with 70% ethanol solution. Proper techniques for working in a flow hood include keeping all utensils/petri dishes towards the back where air is cleanest, always open lids away from you towards the clean air, work in a vertical line of stacked petri dishes so your hands are never positioned over either petri dish/test tube, whatever you want to use. Parafilm works best to seal your growing chambers but saran wrap is a cheap alternative.
The media we work with is the basis of all micro-propagation agars, known as the Murashige and Skoog media, MS for short. Basically they analyzed tobacco leaves, drew a line of best fit through the nutrients found, and transferred it to an agar recipe. A hot plate, preferably with magnetic stirrer, and a pressure cooker will be needed for fix this.
1. Weigh 30 g sucrose
2. Measure out 800 mL double distilled water into 1000mL beaker, combine, this is where a magnetic stirrer/hot plate comes in handy
3. Add 1 mL vitamin B(unsure of exact concentration)
4. Add 10 mL NaFeEDTA
5. Add 20 mL nitration solution
6. Add 10 mL Halide
7. Add 10 mL Myoisotol
8. Add 10 mL Sulfat
9. Add 10 mL PbMO
10. Add BA, cytokinin
11. Add NAA, auxin
12. pH 7.7-7.75, adjusted with varying concentrations of NaOH
13. 8g agar is then added
14. Top to 1000 mL double distilled water
15. Autoclave/pressure cooker for 15 minutes at 20 psi
16. Pour into petri dishes/test tubes, cover and wait to cool.
I realize this isn't a complete formula, I am missing exact concentrations and I forgot how much BA and NAA was added.
As I am sure everyone realizes, the benefits of micro-propagation are immense. I created 6 african violet plants with a leaf about the size of my little thumb and it took less than 5 minutes. Plants have what is known at totipotency, which is basically the ability to form an entire plant with just a single cell of material. It's not an overnight process, the cells must first undifferentiate and form a callus tissue, similar to a tumor. It is from this callus tissue the cells re-differentiate and roots/stems begin to form. Its pretty cool holding entire plants that are sealed in test tubes. I'll post pics of final results tomorrow. I could take a large cannabis leaf and cut in into 75-100 pieces, placed into a few petri dishes, and within a matter of 4-6 weeks have 75-100 newly forming plants. All from 1 leaf.
It really starts getting interesting when they start inserting genes during this process, that's a whole other topic and ethical discussion. I think the US has a hard enough time getting parents to give their children regular vaccines but somehow its O.K. for companies to test bananas on children in Africa that have had the immunizations spliced into the fruit. Or the new corn with Bt inserted into it. I saw a new tomato species that had some desert plants genes inserted into so that it stopped responding to heat and drought stress. Even at like 10 days of 100F+ days, 90F+ nights, healthy and green as could be while the controls were a crisp.
Anyways sorry for the incomplete formula, it should be enough for most of the people actually interested in this to accomplish their goal. Its really not hard, just stay sterile, and use proper techniques. Stay safe/stay medicated-basket
The media we work with is the basis of all micro-propagation agars, known as the Murashige and Skoog media, MS for short. Basically they analyzed tobacco leaves, drew a line of best fit through the nutrients found, and transferred it to an agar recipe. A hot plate, preferably with magnetic stirrer, and a pressure cooker will be needed for fix this.
1. Weigh 30 g sucrose
2. Measure out 800 mL double distilled water into 1000mL beaker, combine, this is where a magnetic stirrer/hot plate comes in handy
3. Add 1 mL vitamin B(unsure of exact concentration)
4. Add 10 mL NaFeEDTA
5. Add 20 mL nitration solution
6. Add 10 mL Halide
7. Add 10 mL Myoisotol
8. Add 10 mL Sulfat
9. Add 10 mL PbMO
10. Add BA, cytokinin
11. Add NAA, auxin
12. pH 7.7-7.75, adjusted with varying concentrations of NaOH
13. 8g agar is then added
14. Top to 1000 mL double distilled water
15. Autoclave/pressure cooker for 15 minutes at 20 psi
16. Pour into petri dishes/test tubes, cover and wait to cool.
I realize this isn't a complete formula, I am missing exact concentrations and I forgot how much BA and NAA was added.
As I am sure everyone realizes, the benefits of micro-propagation are immense. I created 6 african violet plants with a leaf about the size of my little thumb and it took less than 5 minutes. Plants have what is known at totipotency, which is basically the ability to form an entire plant with just a single cell of material. It's not an overnight process, the cells must first undifferentiate and form a callus tissue, similar to a tumor. It is from this callus tissue the cells re-differentiate and roots/stems begin to form. Its pretty cool holding entire plants that are sealed in test tubes. I'll post pics of final results tomorrow. I could take a large cannabis leaf and cut in into 75-100 pieces, placed into a few petri dishes, and within a matter of 4-6 weeks have 75-100 newly forming plants. All from 1 leaf.
It really starts getting interesting when they start inserting genes during this process, that's a whole other topic and ethical discussion. I think the US has a hard enough time getting parents to give their children regular vaccines but somehow its O.K. for companies to test bananas on children in Africa that have had the immunizations spliced into the fruit. Or the new corn with Bt inserted into it. I saw a new tomato species that had some desert plants genes inserted into so that it stopped responding to heat and drought stress. Even at like 10 days of 100F+ days, 90F+ nights, healthy and green as could be while the controls were a crisp.
Anyways sorry for the incomplete formula, it should be enough for most of the people actually interested in this to accomplish their goal. Its really not hard, just stay sterile, and use proper techniques. Stay safe/stay medicated-basket
Tissue culture in the Home Kitchen, go here: http://www.omnisterra.com/botany/cp/slides/tc/tc.htm
Get this book: "Plants from Test Tubes" by Lydian Kyte If you are good with Google you can find a free download. Search for Plants_From_Test_Tubes_UNEDITED.pdf (for some reason, when I posted this, there is a space in the word UNEDITED. Leave the space out before searching. When I tried to leave out the space by editing, the space does not appear.)
Everything described in this post can be done inexpensively and fairly easily.
If you get into this, practice sterile techniques. If you have ever grown magic mushrooms, everything will seem very familiar.
Have fun!
Get this book: "Plants from Test Tubes" by Lydian Kyte If you are good with Google you can find a free download. Search for Plants_From_Test_Tubes_UNEDITED.pdf (for some reason, when I posted this, there is a space in the word UNEDITED. Leave the space out before searching. When I tried to leave out the space by editing, the space does not appear.)
Everything described in this post can be done inexpensively and fairly easily.
If you get into this, practice sterile techniques. If you have ever grown magic mushrooms, everything will seem very familiar.
Have fun!
All this talk about law being in the way of what we do is bollocks..
There is no law to say it cant be done, there are no plants ever needed involved in a production room.
If they are sold as a souvenir or alternatively directly to a licenced company for production with Authorization from the home office.
Or anywhere that there is no cannabis law.
It just means that the novelty to be able to do it ain't going to make millions and its not practical for the un-licensed un-regulated cannabis seed industry.
Has somebody actually then placed stem cell into a artificial seed case and then germinated the "Emblings" as freely and easily as f1 hybrid seeds.
Prove it......
A seed is a ripened and fertile ovule containing an embryonic plant, the only difference in law is that these cultures don't happen in a plants ovary and this can be seen as GM.
Plant patent and seed patent laws brought in after Luther Burbank died has made further issues to owning a strong patent and holding it under international licence without being bullied out of production rights.
Look into actually patenting what genetics your working with first or its game over.
There is no law to say it cant be done, there are no plants ever needed involved in a production room.
If they are sold as a souvenir or alternatively directly to a licenced company for production with Authorization from the home office.
Or anywhere that there is no cannabis law.
It just means that the novelty to be able to do it ain't going to make millions and its not practical for the un-licensed un-regulated cannabis seed industry.
Has somebody actually then placed stem cell into a artificial seed case and then germinated the "Emblings" as freely and easily as f1 hybrid seeds.
Prove it......
A seed is a ripened and fertile ovule containing an embryonic plant, the only difference in law is that these cultures don't happen in a plants ovary and this can be seen as GM.
Plant patent and seed patent laws brought in after Luther Burbank died has made further issues to owning a strong patent and holding it under international licence without being bullied out of production rights.
Look into actually patenting what genetics your working with first or its game over.
S
swisscheese
Bump. Anyone come up with any concrete recipes for cannabis tc yet? I am looking to tc all my strains and already have a hepa flow hood and a dedicated sealed room available. I have limited mycology experience and am a legal medical patient who would be willing to document everything to help others out and I have access to whatever chemicals I need. Really looking for a mentor through this whole process.
