cannabis tissue culture journal

[oxanaca]

so im finally in a position where im ready to start micropropagating. my car broke down and i cant really go anywhere. its cold as hell. and i have everything i need to do it.

ill start out the thread by giving links to all the good research papers, including a paper on somatic embryogenisis (clone seeds)

this one is on hemp it was done by the chinese, their results were inferior to what mississippi was able to do but im going to try their recipe anyways. they have a seed disinfection protocol but it uses mercuric chloride(mercury) which im not messing with. fuckin chinese :rolleyes:
http://www.pakbs.org/pjbot/PDFs/41(2)/PJB41(2)603.pdf

these guys have it going on. im going to put most of my eggs in their basket.
http://home.olemiss.edu/~suman/Directshootorganogenesis.pdf

this outlines a method to successfully produce clone seeds
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3550375/pdf/12298_2009_Article_8.pdf

 

[oxanaca]

the reason i posted this thread was to find others willing and able to start micropropagating right away. ive spend a lot of time reasearching and have suceessfully micropropagated ghost peppers and african violet plants in great numbers. now its time to work on my target plant.

im willing to send 100 milligrams of Thidiazuron, IBA, NAA, Kinetin, GA3, IAA, and enough KOH to dissolve it all. to anyone who builds a transfer area, gathers the nessisary equiptment and seems confident enough to help me reach my goal.

i dont feel like answering any noob questions instead i point anyone intersted to
http://www.amazon.com/Plants-Test-Tubes-Introduction-Micropropogation/dp/1604692065
http://www.amazon.com/Marijuana-Test-Tubes-Micropropagating-Cannabis-ebook/dp/B00FK133XU/ref=sr_1_1?s=books&ie=UTF8&qid=1392784894&sr=1-1&keywords=marijuana from test tubes

one last note before i log off dont anyone buy a kit of any sort, or ill bitch slap you. ill put together a materials list of what i belive is needed for success. i will also be building a culture rotator so i can work with liquid cultures

 

[leadsled]

Tuned in for the show.

 

[madmaster420]

haha bitch slap. what a movie

 

[PButter]

Very neat. Sitting cross-legged with notebook and pencil...

 

[madmaster420]

Very neat. Sitting cross-legged with notebook and pencil...

taking notes on swayze's little bro, no doubt...:p

 

[justanotherbozo]

...'i'll be back', as the saying goes, ...but first i need to bone up on the basics.

thanks for sharin', i'm always open to new ideas, ...new to me anyway, lol.

peace, bozo

 

[audiohi]

Thanks bro! I've been very interested in this also

 

[CBDEMON]

Been waiting for a thread on micropropagation on cannabis!!! hell yeah. Definitely very interested in learning this technique for future projects. I'm on board for this show for sure.

 

[madmaster420]

the reason i posted this thread was to find others willing and able to start micropropagating right away. ive spend a lot of time reasearching and have suceessfully micropropagated ghost peppers and african violet plants in great numbers. now its time to work on my target plant.

im willing to send 100 milligrams of Thidiazuron, IBA, NAA, Kinetin, GA3, IAA, and enough KOH to dissolve it all. to anyone who builds a transfer area, gathers the nessisary equiptment and seems confident enough to help me reach my goal.

i dont feel like answering any noob questions instead i point anyone intersted to
http://www.amazon.com/Plants-Test-Tubes-Introduction-Micropropogation/dp/1604692065
http://www.amazon.com/Marijuana-Test-Tubes-Micropropagating-Cannabis-ebook/dp/B00FK133XU/ref=sr_1_1?s=books&ie=UTF8&qid=1392784894&sr=1-1&keywords=marijuana from test tubes

one last note before i log off dont anyone buy a kit of any sort, or ill bitch slap you. ill put together a materials list of what i belive is needed for success. i will also be building a culture rotator so i can work with liquid cultures

What is the benefit of micropropagation? Hate to sound like a noob, but seems like a lot of extra work?

 

[leadsled]

What is the benefit of micropropagation? Hate to sound like a noob, but seems like a lot of extra work?

Read up

 

[madmaster420]

Read up

okay. will do

bitchslap. hahaha

 

[green bastard]

i dont understand alot about tissue culture so im gonna be following you very close on this one ox

 

[monkeymun]

I am so in for this one. Would love to join you but still don't have the equipment. One day perhaps...

Hadn't you actually started micropropation not so long ago, or were you just researching it? Going to be good to see what you can do with it dude.

 

[oxanaca]

I am so in for this one. Would love to join you but still don't have the equipment. One day perhaps...

Hadn't you actually started micropropation not so long ago, or were you just researching it? Going to be good to see what you can do with it dude.

i havent done it with cannabis just bhut jolokia(ghost peppers) and african violets. i think i may start the process tomarrow tonight i must clean my transfer tent tonight and let the alcohol and bleach air out, and also plan out everything that i will do.

is it just me or do others find that when you type fast on thc farmer it goes haywire and i have to go back and fix things that i typed correctly

 

[ralphy]

Tissue culture really takes growing plants to a whole different level. Hope it all goes well for you, it strikes me as challenging.

 

[oxanaca]

so about 4 days ago i took 20 or so 4 inch long cuttings i added them to 1 gallon of water with 1 ml or physan 20. no phytotoxicity is present at this point. i think im going to follow up by cleaning them ultrasonically for 16 minutes in sterille RO water, then i will transfer them to a 15% bleach solution+ 0.1%tween 20 contained to a side arm flask. i will use a vacuum pump to bring the contents down to a 25mm Hg vacuum. their will be a stir bar in the bottom and i will stir them on a stri plate for 20 minutes. finally i will move the flask to my transfer hood. inside the hood i will break the vacuum and strain them. and rinse them 3 times in sterille RO water, for 5 minutes each.

at this point i will strain them one more time and set them on a sterille napkin. i will cut them down into 1 and 2 node explants and using my massive forceps putt hem in screw top culture vessels containing per liter:

30 grams sucrose(table sugar)
10, 8 or 6 grams agar
full strength MS
ph 5.7
0.11 mg Thidiazuron (0.5 microMoles)

i will then watch. heres where i will be working



here is what i will be wearing

 

[ftwendy]

Rooting for you man. Thanks for sharing.

 

[We Solidarity]

awesome flow hood. Been trying to design one for a while, was it simple?

 

[audiohi]

Very nice DIY cleanroom! I REALLY like it! Where did you locate that tent if you don't mind sharing? Also, what was the cost of your filter for the flow hood? I'm impressed. You seem to have really nailed a cheap workspace!