cannabis tissue culture journal

[oxanaca]

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

its beyond simple, i recommend quality wood for the box though. just do it man

 

[oxanaca]

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

its not as challenging as it appears. i suppose i should do it before i say that though

 

[oxanaca]

it sure was thoughtful of the national institute of drug abuse to fund a study on how to mass multiply cannabis plants
dont you agree? now lets all try not to abuse this knowledge ok

 

[oxanaca]

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!

i bought the tent at a local mega hardware store. look for one early spring.

the filter can be found here, mine was $123, dont be a tool and order a 95% filter its gotta be a 99.99% filter or youll be sorry
http://www.filtera-b2b.com/businessfilters/hepa.htm

99.99% HEPA - Wood Frame - Downstream Gasket - No Header
18 x 24 x 5-7/8 (99.99% HEPA wood) $123.66

you can use an 8 inch inline fan with a speed controller. you must have a plenum the same size as the filter after the fan. you also need a prefilter. the trick to finding the correct air speed is to hold a lighter in front of the filter and adjust the speed so that the flame bends over but doesnt blow out. as an alternative you can use a manometer with a hose connected to your plenum and create 1 inch or pressure on a water column.

 

[audiohi]

Word.... Thanks for the tips! 99.99% is the only way to go... I drew up a large one for mycology.... but never followed through due to the cost. Right back to the stupid glove box I went. Yours looks very nice with a great price. Do you get the sideways lighter flame with that one? 100ft/sec if I remember correctly? Maybe 1ft/sec? I barely even remember anymore..

 

[oxanaca]

Word.... Thanks for the tips! 99.99% is the only way to go... I drew up a large one for mycology.... but never followed through due to the cost. Right back to the stupid glove box I went. Yours looks very nice with a great price. Do you get the sideways lighter flame with that one? 100ft/sec if I remember correctly? Maybe 1ft/sec? I barely even remember anymore..

yeah i have to turn it down to the 75% mark on the controller

100ft/second would blow you away thats 68mph. its actually 100feet/min or a little over 1 mph

 

[oxanaca]

came up with an interesting idea. i dissolved some agar in some water on a flying pan on the stove and stuck some stems in it with a forceps. unfortunately i didnt measure how much i added and it come out perfect. they stuck in easily but were still supported, so unlike MEA agar for fungi. tomorrow i will test 9 different concentrations in the 5 to 10 g/l range. and see if i cant find the minimum concentration that will adequately support the explants, while remembering that the plants will get quite large as the shoots grow out and may fall over.

i had a tough time with the small ones, forceps i was using was to big for the job. mississippi university used 1cm long explants im going to stick with that and try a few 2 and 3 node explants as well

 

[oxanaca]

this guy had some success, ive never seen a picure of roots invitro in a forum before
http://buymarijuanaseeds.com/community/threads/tissue-culture.149980/

looks like they had to firm of a gel at 10g/l agarose
i wonder why they did that agarrose has a higher gel strenght than regular agar. no wonder they lost so many roots

 

[monkeymun]

What's the idea behind the agar+stems?

 

[oxanaca]

i was just curious how it would stick into the agar

 

[oxanaca]

opps remember where i said i left cuttings in physan 20 solution in the fridge for 4-5 days. dont do that. on a flat of conventional cuttings i stuck im seeing phytotoxictiy on 10% of the new growth 3 days after the soak. i guess it takes a while for the damage to show. im going to think this threw a little more before i take another step.

 

[iCultivate]

You're like a mad scientist Oxacana!

Other than doing this for the sake of it, what do you hope to achieve?

-- iCultivate --

 

[oxanaca]

You're like a mad scientist Oxacana!

Other than doing this for the sake of it, what do you hope to achieve?

-- iCultivate --

hardly i just have a good memory and lots of energy and drive

+ disease and bug free starts

+ extra bud in place of the mothers i would have to light using the traditional propagation method.
the way i see it if i discontinue 1000 watts of HID lighting being used to illuminate mothers 18 hours a day. i will be able to redelagate that energy to 1500 watts of light illuminating flowering plants and score an extra 750g's on average every month, once i get things down pat.

+ i want to have 100+ kinds of weed at my finger tips.

+ mass multiplication

+i want to select and maintain elite autoflowering genetics so i can grow them outside during june-august. 2 month crop cycles when the sun is the most intense, and nobodys looking for it.

+clone seeds. once i have an awesome portfolio, i want to be able to maintain it even if i move to a new country, i will be able to have someone mail me envelopes with clone seeds that i created and immediately start up again with my trusty elite plants.

 

[iCultivate]

Good list of ambitions! Very keen to see if you can achieve these once you've mastered this science.

-- iCultivate --

 

[oxanaca]

practiced what im about to do. i made up several different concentrations of agar all ph'd to 5.7. i tryed 2, 4, 4.5, 5, 5.5 6, 8 10, and 32 grams per liter scaled down to 250ml. i found that 4 grams per liter didnt gel properly and 7 grams per liter was too hard. between 4.5 and 6 grams seemed right. im going to run 5 grmas per liter in my intial experiments.
using this agar, unfortuantly lost my good micropropagtion agar and gellan gum,i ll find it though
http://www.amazon.com/Telephone-Brand-Agar-Agar-Powder/dp/B007PQTVSY

turns out i lied i am a mad scientist muhahaha

im going to use baby food jars with magenta b caps. ole miss says 1 cm cuttings works best so thats what im going to try
stay tuned

 

[Delerium]

I'm liking this, had 25 varieties last year that I had to chuck cuz my mom room was getting too crowded.
I was going to get one of those kits from http://planttc.com, I just didn't know if it was a waste or not. But for a beginner it might be worth a shot?

 

[oxanaca]

it doesnt include thidiazuron, which you need for cannabis. its just a bunch of marked up diluted junk. i purchased a kit from them. they made me buy a membership or some shit it was like $40 and i got like 3 little bottles of hormones with 10mg of each hormone diluted in each.

 

[Herb Forester]

Nice project. In case you haven't seen, here's another article you might like.
http://skunkpharmresearch.com/tissue-culture/

 

[oxanaca]

Nice project. In case you haven't seen, here's another article you might like.
http://skunkpharmresearch.com/tissue-culture/

yep there following the same protocol as me

 

[oxanaca]

i did a dry run of the tissue culture disinfestation protocol, and practiced asepticaly transfering explants to media. i really want to knock it out of the park on the first try






tomarow night im going to transfer 25 disinfested explants to baby food jars. ill post how it went