Or_Gro
Well-Known Member
Yep.
Or_Gro’s Tissue Culture Journal
- Thread starter Or_Gro
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- tissue culture
Or_Gro
Well-Known Member
Gettin there....
Gonna test out a less expensive “air purifier” as a substitute for my hepa-filtered laminar-flow fan.
It is “true hepa”, which is a marketing phrase....all hepa filters, by definition, can filter out particles down to 0.3 micron, removing 99.97% of particles.
I got the best pick, Coway purifier, in the following article:
https://www.google.com/amp/s/thewirecutter.com/reviews/best-air-purifier/amp/
The idea is to find a small, draft-free area, close it off, sanitize working surfaces, and run air-purifier until air is “clean”; Then do tissue culture work using sterile technique.
This first go-round, i’ll do work in a grow tent. The following time, i’ll do work in a bathroom.
I transplanted my cuttings this morning, so a little cleaning and resetting, and I’ll be ready to do the first kit.
Gonna test out a less expensive “air purifier” as a substitute for my hepa-filtered laminar-flow fan.
It is “true hepa”, which is a marketing phrase....all hepa filters, by definition, can filter out particles down to 0.3 micron, removing 99.97% of particles.
I got the best pick, Coway purifier, in the following article:
https://www.google.com/amp/s/thewirecutter.com/reviews/best-air-purifier/amp/
The idea is to find a small, draft-free area, close it off, sanitize working surfaces, and run air-purifier until air is “clean”; Then do tissue culture work using sterile technique.
This first go-round, i’ll do work in a grow tent. The following time, i’ll do work in a bathroom.
I transplanted my cuttings this morning, so a little cleaning and resetting, and I’ll be ready to do the first kit.
led1k
Well-Known Member
I feel like a kid at Christmas I’m so psyched to see your process and results!
Or_Gro
Well-Known Member
Here’s the victim:
Amnesia OG, aka Where’s My Bike
Here’s the lab:
Clockwise from left in this 4x4:
Hepa filter (blue=clean), lab table/cloning tote, home depot lab chair. Have wiped them down in 70% iso; hepa running, tent zipped.
Still packing my convertible supplies tote/ open-front glovebox:
Or_Gro
Well-Known Member
Ok, almost there...
Decided, wtf, i have a sterilizer/pressure cooker, why not use it?
Pressure cooker (PC) sterilizing process:
1. Get all your sh!t together, and prepare.
2. Fill PC with water, to appropriate level, insert wire rack/trivet to keep contents out of water.
3. Add ~10mL (~3/4-1” depth) nutrient solution in each test tube, place cap on tubes but do not tighten (unless you want a f’n mess), put tray with nutrient tubes in PC.
4. Loosely wrap scalpel, forceps, scissors, paper towels in aluminum foil, and place in PC.
5. Put cutting plate in PC.
6. Put lid on PC, tighten appropriately, open steam valve, turn on heat.
7. When steam starts coming out of valve (mine takes about 50 mins to get up to steaming speed), let run for 5 minutes, then close steam valve/put on steam weight, when pressure builds to 15psi (mine takes about 45 minutes to build), run for 20 mins.
8. After running for 20 mins @15psi, shut off heat. Let cool to room temp (overnight), DO NOT OPEN until you have moved pressure cooker into your sanitized “lab”.
9. When ready to do your tissue work, open PC, arrange items for good workflow.
Decided, wtf, i have a sterilizer/pressure cooker, why not use it?
Pressure cooker (PC) sterilizing process:
1. Get all your sh!t together, and prepare.
2. Fill PC with water, to appropriate level, insert wire rack/trivet to keep contents out of water.
3. Add ~10mL (~3/4-1” depth) nutrient solution in each test tube, place cap on tubes but do not tighten (unless you want a f’n mess), put tray with nutrient tubes in PC.
4. Loosely wrap scalpel, forceps, scissors, paper towels in aluminum foil, and place in PC.
5. Put cutting plate in PC.
6. Put lid on PC, tighten appropriately, open steam valve, turn on heat.
7. When steam starts coming out of valve (mine takes about 50 mins to get up to steaming speed), let run for 5 minutes, then close steam valve/put on steam weight, when pressure builds to 15psi (mine takes about 45 minutes to build), run for 20 mins.
8. After running for 20 mins @15psi, shut off heat. Let cool to room temp (overnight), DO NOT OPEN until you have moved pressure cooker into your sanitized “lab”.
9. When ready to do your tissue work, open PC, arrange items for good workflow.
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schmebulock
Well-Known Member
seems like this is all the same shit you gotta do to grow shrooms lol
i'm sure putting in all these steps and effort will pay off with a wonderfully sterile environment!
Or_Gro
Well-Known Member
Yep, if you know how to do shroom in vitro culture, you’ve already got the sterile tek part nailed....
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schmebulock
Well-Known Member
the video was interesting to watch. so am i understanding it correctly that this just enables you an even quicker route to a mature veg plant ready for flower than standard cloning?
edit: without the need for a mother plant too
edit: without the need for a mother plant too
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Or_Gro
Well-Known Member
As i said earlier...
No, this just enables you to take a cutting and prep it for rooting in a plug in 8-12 weeks...
I’m doing it to introduce in vitro sterile work. Knowing how to do sterile tek competently is the main skill involved in any in vitro process.
The real benefit of tissue culture for cannabis comes from the in vitro multiplication/replication and rooting stages, that i will be doing in the next kit....
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schmebulock
Well-Known Member
lol man you're on a whole 'nother level. fascinated to see how this goes.
Or_Gro
Well-Known Member
This is actually simple sh!t, you just gotta do it right, which is just another set of skills to learn.
Plants have a lot of undifferentiated cells that will become specific tissue when exposed to certain hormones.
Using this, we can create miniature plants, almost suspended in time; that can be transplanted from testtubes/jars, in 1:multi-hundred replications, to individual buckets/pots; which can become full-sized duplicates of the mother plant, and mothers themselves.
[Imagine ordering a desired strain/pheno, and a couple days later a delivery truck drops off a plant....]
Just a matter of keeping the cells alive (w sugar & water/agar & appropriate conditions), keeping the microbes out of the sugar (sterile tek), and applying the correct hormone & rooting (next kit).
‘Be great to have a use for my expensive supercloset supply cabinet again. Pack it with all my fave phenos. Pull em out and make copies to grow or trade...
Plants have a lot of undifferentiated cells that will become specific tissue when exposed to certain hormones.
Using this, we can create miniature plants, almost suspended in time; that can be transplanted from testtubes/jars, in 1:multi-hundred replications, to individual buckets/pots; which can become full-sized duplicates of the mother plant, and mothers themselves.
[Imagine ordering a desired strain/pheno, and a couple days later a delivery truck drops off a plant....]
Just a matter of keeping the cells alive (w sugar & water/agar & appropriate conditions), keeping the microbes out of the sugar (sterile tek), and applying the correct hormone & rooting (next kit).
‘Be great to have a use for my expensive supercloset supply cabinet again. Pack it with all my fave phenos. Pull em out and make copies to grow or trade...
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schmebulock
Well-Known Member
TIME TO SCIENCE!
Or_Gro
Well-Known Member
led1k
Well-Known Member
Love the delivery truck idea but for now your last paragraph is why I'm doing this for sure! One rack of
tubes space wise and 36 of your favorites? Disease free?
Or_Gro
Well-Known Member
The process:
Sample material soaking 30mins in 500ppm chlorine solution
Sample material moved inside “hood”, soaking in 500ppm chlorine, cutting plate w pool of 500ppm chlorine solution
Sample material with big leaves removed (done when cutting taken; 1/4” stub left of each petiole during 30 min soak)
Petiole stubs removed; stem end removed, cut to length so that first node from bottom is above surface of nutrient solution; leaves cut off/smaller
Sample placed in tube, capped, sealed w breathable film, labeled w contents and date
1st node above surface of nutrient solution
Tissue work completed
Their new home in a 2x3
Under a single 96elite, runnin
Sample material soaking 30mins in 500ppm chlorine solution
Sample material moved inside “hood”, soaking in 500ppm chlorine, cutting plate w pool of 500ppm chlorine solution
Sample material with big leaves removed (done when cutting taken; 1/4” stub left of each petiole during 30 min soak)
Petiole stubs removed; stem end removed, cut to length so that first node from bottom is above surface of nutrient solution; leaves cut off/smaller
Sample placed in tube, capped, sealed w breathable film, labeled w contents and date
1st node above surface of nutrient solution
Tissue work completed
Their new home in a 2x3
Under a single 96elite, runnin
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Smokexbreak
Well-Known Member
Holy fuck this tripped me out!