I just did the opposite...thinking f* it. only live once.... may as well drink Glenfiddich...well unless I figure out how to transfer my mind to this 15 year old clone I have in the tank.
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Cannabis tissue culture
- Thread starter Gray Wolf
- Start date
I just did the opposite...thinking f* it. only live once.... may as well drink Glenfiddich...well unless I figure out how to transfer my mind to this 15 year old clone I have in the tank.
it would still be only whatever the plant material dried weighed..so 10 tissue culture samples dried would be like what less then an 1/8th? i don't see that being a huge deal with domestic distribution. international and customs may be a different story though.
use ebay for flowhoods/hepa's. one can set up a full(non pro) lab for around 1k room permitting.
good site that aggregates ebay hepa's:
http://www.fungifun.org/English/Flowhood
I picked up several 2'x2'x10" last year for $50 a piece.
good site that aggregates ebay hepa's:
http://www.fungifun.org/English/Flowhood
I picked up several 2'x2'x10" last year for $50 a piece.
smokingjoe_
Member
I purchased a home based TC kit from a supplier via eBay which cost me about $40 with the intention of using it to preserve desirable phenos, but have never got around to putting it to use. I'd really love to build a sealed clean room when we build new digs in a few years time.
To be honest the only thing that stopped me was my inability to settle for an upturned box as a hood and difficulty justifying the expense of a laminar flow hood to the commissioner of fun and finance.
Chimera those Frogs eggs certainly sound interesting, do you have a post relating to them?
To be honest the only thing that stopped me was my inability to settle for an upturned box as a hood and difficulty justifying the expense of a laminar flow hood to the commissioner of fun and finance.
Chimera those Frogs eggs certainly sound interesting, do you have a post relating to them?
Did my first Cannabis TC more then 20 years ago after I self taught myself and bought all the needed lab equipment and set up a small in-vitro lab.
I found it pretty easy to Clone in-vitro meristems, Callus even easier, but I did have trouble getting the Callus to shoot after rooting. Rooting is easy.
Growing meristems and Callus is easy, so is sterilizing a clone so you can stick it in agar and nutrients to grow.
Give it a try, it is easy, and if anyone figures out how to freeze Callus or Meristems and then thaw them out and re-grow them please let me know, that was my real interest in the first place. When I did it they were reduced to black slime lumps, zero life...
As for freezing, I had a professional In-vitro Company try and find a way to freeze meristems or Callus, but after a few years gave up, I still think it will be done by pretreating the Callus or Meristems or other newer protocols in use today that we did not try.
My dream was a library that would maintain varieties or unique individuals in a very small space, for very long times, that could be re-grown if needed. Keeping clones alive is such a pain and waste of time, space and energy. Freezing would be great!!
-SamS
I found it pretty easy to Clone in-vitro meristems, Callus even easier, but I did have trouble getting the Callus to shoot after rooting. Rooting is easy.
Growing meristems and Callus is easy, so is sterilizing a clone so you can stick it in agar and nutrients to grow.
Give it a try, it is easy, and if anyone figures out how to freeze Callus or Meristems and then thaw them out and re-grow them please let me know, that was my real interest in the first place. When I did it they were reduced to black slime lumps, zero life...
As for freezing, I had a professional In-vitro Company try and find a way to freeze meristems or Callus, but after a few years gave up, I still think it will be done by pretreating the Callus or Meristems or other newer protocols in use today that we did not try.
My dream was a library that would maintain varieties or unique individuals in a very small space, for very long times, that could be re-grown if needed. Keeping clones alive is such a pain and waste of time, space and energy. Freezing would be great!!
-SamS
Very interesting stuff here. Ive been fascinated with the idea of housing 100s of genotypes, maintenance free, taking up little space, and not having to devote an entire room to house them. Ive seen those home kits for ~$200, are these all I would need to get started? they sell kits for african violets, would these kits contain the same hormones and things I would need to take callous cultures from cannabis
smokingjoe_
Member
The Revolution, as I understand it (and that's not widely) different cells will react to different hormones depending on concentration and type, and it's all a bit of trial and error.
eBay tissue culture kit and you'll get a few hits
eBay tissue culture kit and you'll get a few hits
Did they try the standard PEG/DMSO? I think this method came into common use in the mid '80s- if your efforts were before this, I would try it, I assume that it is pretty effective for most plants. I think they sometimes create a suspension of callus cells and freeze that, I've heard that this helps freeze the cells very quickly, which is important.
Patent deposits of TC at ATTC are required to be frozen callus, so it must be the standard way of preserving TC?
I know it works really well for beer yeast cultures.
PEG/DMSO was tried, with no luck.
Although Cannabis seeds or pollen can be frozen with no negative effects, they still grow fine, Cannabis plants and roots don't survive freezing.
Some plant cells take freezing with no treatment, some like Cannabis are very hard or impossible so far to freeze, thaw and re-grow. Even if eventually successful then it needs to be determined if freezing causes mutations or not? If it does not cause mutations then freezing will be a useful tool for keeping long term collections of germ-plasm alive in a cheap easy small space. I was mostly trying freezing meristems not callus, as at the time, 20+ years ago, getting callus to shoot was not so easy, so I focused on meristems, which although I presumed are even harder to freeze and thaw, and re-grow, but meristems are real easy to grow and reproduce in-vitro harden off, and plant out to grow and flower. But in the end neither callus or meristems could be frozen thawed and re-grown, so I gave up.
-SamS
Although Cannabis seeds or pollen can be frozen with no negative effects, they still grow fine, Cannabis plants and roots don't survive freezing.
Some plant cells take freezing with no treatment, some like Cannabis are very hard or impossible so far to freeze, thaw and re-grow. Even if eventually successful then it needs to be determined if freezing causes mutations or not? If it does not cause mutations then freezing will be a useful tool for keeping long term collections of germ-plasm alive in a cheap easy small space. I was mostly trying freezing meristems not callus, as at the time, 20+ years ago, getting callus to shoot was not so easy, so I focused on meristems, which although I presumed are even harder to freeze and thaw, and re-grow, but meristems are real easy to grow and reproduce in-vitro harden off, and plant out to grow and flower. But in the end neither callus or meristems could be frozen thawed and re-grown, so I gave up.
-SamS
Wow, sounds like a worthy challenge! At this point, we are just trying to do the full tissue culture cycle to adult plant, but will pick up on freezing as a followup project. Sounds like it isn't easy or casual!
Keeping any number of strain cultures going at this point seems simple, but even in clean rooms, contamination spoors usually eventually find their way in. Keeping a library of strains viable may prove a challenge, even without addressing cryogenic suspension!
C
chefro420
~deadwood~
New member
Sam, Could you be so kind as to tell us what you used for shoot differentiation from callus? I have been doing some research, but have yet to find a starting point where to begin my attempts with getting shoots from callus. I know that the whole process is less than reliable process, but something I am very interested.
You do not want to use what I did as I had very very low rates of shooting, like less then 5%. You will find newer articles with formulas for shooting, you will find rooting easy, no problem. Maybe someone else can help? Check the Italians they work with in-vitro Cannabis a bunch, and have told me they get more then 10% to shoot.
-SamS
-SamS
~deadwood~
New member
If anyone knows of any published studies or threads talking about shoot differentiation from callus please point me in the right direction. My google-fu is weak it seems.
A note on viruses and this kind of culture. While the virus levels in the meristem are generally extremely low to non-existent, they're not always completely absent. However you can clear most viruses through a few rounds of high-temp cultures. Never did anything seriously with culture though.
I know there was some work years ago into using antifreezes, sorry "cryoprotectants" combined with rapid chilling & a final immersion in liquid nitrogen to preserve callus, no idea how it panned out.
I know there was some work years ago into using antifreezes, sorry "cryoprotectants" combined with rapid chilling & a final immersion in liquid nitrogen to preserve callus, no idea how it panned out.
Yes we do!
Progress is being made! GG moved the first batch to the rooting solution and it is looking robust.
Here is a picture of Red Diesel and of a Penicillin contaminated U of Dub. We also lost one to bread mold.
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wrongwrong
Member
Sharing is caring
I'd love to see your babies get around
Haha you want him to impregnate the women of the world insted of spending his time with canna.... thats taking sharing to a new level! haha
