cannabis tissue culture journal

  • Thread starter oxanaca
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oxanaca

oxanaca

322
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Dont anyone buy magenta b caps made by phyto technology. They dont come off smoothly. You need the real magenta brand.

Also my seedlings havent been put under any light(hence the leggyness. Im going to do that now with the healthy ones. And hopefully start a new batch of seeds, Tomarrow using what ive learned so far.
 
oxanaca

oxanaca

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So im ready to go another round. I have decided to devote and desighn an entire room around sterile culture. Its a 11x12 room. Heres a diagram
20150109 113226

, all walls will be covered In 3 mil poly. The floor will be a sheet of 3 mil black poly. With a sheet of 0.7mil plastic that will be replaced frequently. Havent decided what to use for the partition walls.
 
oxanaca

oxanaca

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In addition to the walls ceiling and floor bing covered in plastc. I will also have two foot baths before the aseptic transfer room. You must go thru 3 doors to get to the transfer room.

The room will be positive pressure. This will happen by pulling air from outside the room and running it thru an initial hepa hilter and then my laminar flow hood. I will keep my hepa system running 24/7

I will run ozone threw my air handeling system a couple hours a week. I will also find a way to fog disinfectants to keep all surfaces clean. The lab will be thoroughly cleaned weekly. Everything brought into the room will be wiped down with 70 iso.

Gonna get a real autoclave to. So I dont have to struggle carrying a dirty pressure cooker from the kitchen
http://www.sterilizerautoclave.com/all-american-75x-electric-sterilizer-013
 
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oxanaca

oxanaca

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63
I've decided to start all seeds in 50 milliliter centrifuge tubes. I will then switch to baby food jars for two more transfers. And if I have a low contamination rate I will go to my magenta 3x3x3 ga7 culture vessels.
 
oxanaca

oxanaca

322
63
So another little update. I disinfected 25 seeds. Using this method.

24 minutes in an ultrasonic cleaner with 3% h2o2 and 0.1% tween 20

30 second dip in 70% isopropanol

Rinsed in sterile distilled water

20 minutes in 10% chlorox bleach + 0.1% tween 20 solution.

Rinsed with sterile distilled water

Aseptically transfered to

20 baby food jars containing per liter
10g sucrose
7g telephone brand agar agar
1/2 ms
ph5.7

5 baby food jars containing coarse vermiculite hydrated to field capacity.

20150128 031302
 
oxanaca

oxanaca

322
63
So ive sort of hit a wall. I want to test 7 or 8 different seed sterilization protocols out to find one that kills no seeds and leads to no contamination. Problem is I need about 300 seeds to do it. I have about 50 good looking seeds ive collected. but thats no where close to what I need. And I dont know where to buy weed with seeds in it.

I know theres someone that has some balls and a pile of viable mexican seeds. They are looking to part with. Im not asking for quality seeds just sproutable seeds.

So if you can help, shoot me a pm and we can get the cannabis tissue culture really cranking. Without any wasted $10 seeds.

to science
 
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GrowingINheaven

GrowingINheaven

84
33
I have experience in micropropigation of orchids And potatoes. I started a few hundred test tubes, my dog ended up knocking over the shelving they were on. I Think I still have all my materials. I'll post up why I have in a day or two when I have time to hit up storage and grab everything. Much luv.
 
GrowingINheaven

GrowingINheaven

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33
oxanaca said:
So ive sort of hit a wall. I want to test 7 or 8 different seed sterilization protocols out to find one that kills no seeds and leads to no contamination. Problem is I need about 300 seeds to do it. I have about 50 good looking seeds ive collected. but thats no where close to what I need. And I dont know where to buy weed with seeds in it.

I know theres someone that has some balls and a pile of viable mexican seeds. They are looking to part with. Im not asking for quality seeds just sproutable seeds.

So if you can help, shoot me a pm and we can get the cannabis tissue culture really cranking. Without any wasted $10 seeds.

to science
Click to expand...
Pm me, I gotch ya
 
O

Oldchuck

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@oxanaca, there is an easy solution to your seed shortage problem. Hemp seed. Same species as your $10 per seed stuff. I bought several thousand seeds for $20, ordered from Finland, about a pound of seeds for endless experimentation. I also bought an ounce of viable hemp seed from California. And to top it off, I bought 40 grams of hemp buds from Slovenia ordered through Amazon of all places. Nice buds, lots of seeds, totally worthless for either medical or recreational purposes. But for your experiments I would think those would work fine.
 
oxanaca

oxanaca

322
63
Oldchuck said:
@oxanaca, there is an easy solution to your seed shortage problem. Hemp seed. Same species as your $10 per seed stuff. I bought several thousand seeds for $20, ordered from Finland, about a pound of seeds for endless experimentation. I also bought an ounce of viable hemp seed from California. And to top it off, I bought 40 grams of hemp buds from Slovenia ordered through Amazon of all places. Nice buds, lots of seeds, totally worthless for either medical or recreational purposes. But for your experiments I would think those would work fine.
Click to expand...

Wow thats an awsome idea. I never would have thought about hemp. Ill start looking right away.

What does a hemp bud look like lol.
 
O

Oldchuck

27
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Yes, for seizure control.

Finola hemp works very well. Slovenian hemp doesn't work at all.
 
CPT Dann

CPT Dann

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3
OH man, I wished this thread had continued. Is there anyone out there that has continued with TC?
 
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