cannabis tissue culture journal

[audiohi]

Are you still working on this project oxanaca?

 

[ralphy]

Would like to know the answer to that also.

 

[buffbuds]

Hey I'm super interested in all of this, I have everything I need already to go, all the baby food jars, magenta b-lids, Iaa, Naa, B(ap), and others ready to go. I have to get back to you, but I'm doing agar with 900 g/m strength, and have had limited sucess, but with promising results. My sterility is fine, being a brewer I understand the game already, and have supplies to keep myself happy and clean. I will be building a multi purpose room with laminar flow hood soon, but basically, I'm stoked to find someone else doing this, and am excited to get back into it myself.

Questions

Whats the lighting requirement change over the different stages of genesis and growth?
I have a 2x4x7 tent I'm going to build up with racks and lights, but wanted input about that, are wire racks ok? how many 4 foot t-5 bulbs would you recomend me using per rack?

 

[Sativied]

Unfortunately this seems typical for TC journals... starts out with a lot of noise and ends up dead quiet.

 

[ethnoman]

Probably when the person realizes how much work and money TC really is and decides that growing plants in more traditional ways is ultimately more productive more small scale growers.

 

[ralphy]

It does seem that way...

 

[buffbuds]

I am looking to do Tissue culture for an unlimited plant count grow system to make more consistent and vigorous plants with a controlled propagation and the ability to verify sterility of plants through the visors of tissue culture. It least to more vigorous root systems and a more uniform plant. also, its a way to ship samples and viable plants across the world, as there is no regulation of plant tissue samples for "research". so this is the only legal way to send clones of genetics to other US states from a legal state.

 

[ethnoman]

Start learning then! It's a complex and multifaceted technique and high initial capital investment.

 

[RHoG]

I'm gonna jump in here, maybe wake this thread up with my opening post.

On Tissue Culture and Microprobagation in general

If you like the planttc kit, go ahead and get it. I think it's a good single source for starting materials you're gonna need. A little pricey, but good for lazy stoners 'cause it's all in one box. Just know that the kit alone will not get you producing clones in culture.

You're gonna have to do a bunch of self education. If you already have a technical or science education then great, you're ready to go. If not, you will have additional reading to do. I've noticed that even in the Overgrow era, which got us to this point, people that have successfully learned this technique are keeping quiet about the finer details.
(Ya hear me skunkpharm?)

I'll be posting in this thread and we'll see how it goes. If you're parts of the group waiting for good info, here is the major resource;

Most of the research has been done at the University of Mississippi by a guy named
Suman Chandra. All of this work has been published in the relevant peer reviewed journals. Some is freely available as PDF's, some of it in books by same author, some of it you just have to buy. I'll bet someone condenses it in a more readable, un-prosecutable form before to long ;)

I've seen some people opine that these techniques are not that productive, are hard to learn, don't really work, and so on.

While some of that might be true, don't doubt the ability of these new techniques to massively multiply your source stock well beyond what simple cloning could ever do, reawaken tired genes, and protect your precious genotype for the long term..

 

[Sativied]

I'll be posting in this thread and we'll see how it goes.

Will you also actually be doing it, as in some sort of log?

And thanks for pointing out Suman Chandra. You just pulled open a can of candy for me, I'm very interested in TC. My main concern is that I won't be able to set up a proper space to do so.

It would be great if you could start with what is needed besides the TC kit resources, i.e. a sterile tent or something, is that doable?

"Just know that the kit alone will not get you producing clones in culture."
Is that because of the kit not being complete, i.e. need to buy hormones or w/e separately, or as in you obviously need to learn a lot as it doesn't contain a simple step by step?

 

[oxanaca]

I'm gonna jump in here, maybe wake this thread up with my opening post.

On Tissue Culture and Microprobagation in general

If you like the planttc kit, go ahead and get it. I think it's a good single source for starting materials you're gonna need. A little pricey, but good for lazy stoners 'cause it's all in one box. Just know that the kit alone will not get you producing clones in culture.

You're gonna have to do a bunch of self education. If you already have a technical or science education then great, you're ready to go. If not, you will have additional reading to do. I've noticed that even in the Overgrow era, which got us to this point, people that have successfully learned this technique are keeping quiet about the finer details.
(Ya hear me skunkpharm?)

I'll be posting in this thread and we'll see how it goes. If you're parts of the group waiting for good info, here is the major resource;

Most of the research has been done at the University of Mississippi by a guy named
Suman Chandra. All of this work has been published in the relevant peer reviewed journals. Some is freely available as PDF's, some of it in books by same author, some of it you just have to buy. I'll bet someone condenses it in a more readable, un-prosecutable form before to long ;)

I've seen some people opine that these techniques are not that productive, are hard to learn, don't really work, and so on.

While some of that might be true, don't doubt the ability of these new techniques to massively multiply your source stock well beyond what simple cloning could ever do, reawaken tired genes, and protect your precious genotype for the long term..

I was actually going to update this last night believe it or not.
I dropped some seeds, which were disinfected in 3% h2o2 for 4 hours, in baby food jars.
Recipe was 1 liter distilled water, 30g sucrose, ph 5.8.

I forgot to add my ms salts. I also didnt rinse my seeds off after disinfection as I forgot to sterilize my rinse water.

Well have to see what happens. Im going to do another round of disinfection, with seeds and live tissue. Nomarrow

 

[Sativied]

I bookmarked a couple of links a while ago, not sure if they are still current but surely some of the info is:

 

[oxanaca]

I bookmarked a couple of links a while ago, not sure if they are still current but surely some of the info is:

These are good reads but outdated. From what I can tell you really need tdz to be successful.

Heres the best source ive been able to find

Heres a great source for any other hormone u may need

 

[RHoG]

Bill Graham seems like a nice guy, a lab nerd angling to get some of the money that's expected to flow out of legalized medical cannabis. And that's ok because I'm a lab nerd too. And like Mr. Graham, I have my own lab.
Any information I give here assumes that you have the kit. It's not necessary but it gives a common reference, capiche?

I haven't decided yet on exactly how much, in the way of pics and what not, I'll post here. One can't be too careful.

Oxanaca has done a great job so far and I'll add as needed.

So yes, you're gonna have to add some hormones as Oxanaca has indicated.
Read the indicated reference, that's got everything you need and
learn what sterile technique is, and get yourself a work area where you can work accordingly. The number of transfers a successful program requires demands you work cleanly.

When you take ex plants, make sure that there is a tiny little start in the crotch of the selected material... This is what is meant by axilary...

And I wanted to add that the hormones are available in kit form at eBay

Sativied, the "shoot tip" source is good, the lycaeum stuff is not. I don't believe for a second that they got shoot off of callus

 

[oxanaca]

Thought I would add a picture or 2.

16 of 29 seeds have germinated. Things are looking up. I bet I wont need the ions from the ms salts for seed initiation medium. Gonna add ms salts tomarrow though and compare.

Evcited to get this going ive wanted to do this with cannabis since my success with ghost peppers.

 

[oxanaca]

Im noticeing a little bit of fungal growth on some of the seeds. I Know h2o2 isnt 100% effective on vegetative fungi, due to my experiance with peroxygenated agar in the mushroom hobby.

I think im going to add an ultrasonic cycle before the h2o2 bath. And a short 10 % bleach bath after. Before finally rinsing with sterile h2o.

Also 23 of the 29 seeds germinated. So 3% h2o2 obviously. Isnt detrimental to there health. Even at 4 hrs submersion.

 

[Seamaiden]

Would methylene blue or malachite green have a place here? In fishkeeping, fish eggs are stained with malachite blue for two reasons--help show which eggs are not viable (they'll stain blue if they're inviable) AND as an antifungal. It's also used in surgeries where the surgeon needs to easily see what tissue is infected.

 

[oxanaca]

Would methylene blue or malachite green have a place here? In fishkeeping, fish eggs are stained with malachite blue for two reasons--help show which eggs are not viable (they'll stain blue if they're inviable) AND as an antifungal. It's also used in surgeries where the surgeon needs to easily see what tissue is infected.

Wow thats interesting, ill have to look into those. I think eventually I want to spend some time learning about fish, coral. And the science behind that environment. I think it would open my eyes, and be bring me one step closer to becoming a master cannabis grower

 

[lino]

Nice info. I already order my chemicals from a link you provided before I saw this thread. Do you use any different seed germ agar recipes for seed propagation cultures?
I noticed new hormone/chemicals/regulators now avaliable that prompted me to try some new recipes.

 

[oxanaca]

So 8 of 29 seeds grew a black spored mold. No other instances of contamination, it all was from the unclean seeds, my sterile technique was sound, but the 3% h2o2 disinfection treatment wasnt enough. I think I was correct about h2o2 not being effective on live fungal cultures.

6g of telephone brand agar per liter, produced a firm suportive gel. When I pulled a seedling out with a root 1.25inches long the root wasnt damaged. I tested the ph of the finished gel after I heated my gel to dissolve the agar, i waited for the gel to reach 120 degrees (its still a liquid at this temperature).before I took a reading with my hORIBA twin drop pH meter. The pH rose from 5.8 to 7. 3. I will switch to a more professional type of agar to avoid this