Or_Gro’s Tissue Culture Journal

Or_Gro

Or_Gro

Well-Known Member
Or_Gro said:
Beginning day 2:

View attachment 4349752 View attachment 4349750 View attachment 4349749

On contamination watch, 2-3 weeks til ready to root....
Click to expand...
Again: this is an inefficient approach for rooting cannabis, rooting can be done with a lot less foreplay using scissors and an aerocloner in ~10 days....

This method is being demonstrated, to show the sterile technique, and to test effectiveness of a less expensive hepa fan rather than an expensive full-blown laminar-flow setup....

Next up, real tissue culture: taking slices of these samples, turning them into miniature plants, rooting them in tube/jar, then storing some plants and transplanting others to normal growing environment.
 
led1k

led1k

Well-Known Member
Or_Gro said:
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.

View attachment 4348870

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.

View attachment 4348875

4. Loosely wrap scalpel, forceps, scissors, paper towels in aluminum foil, and place in PC.

View attachment 4348871 View attachment 4348872 View attachment 4348873 View attachment 4348874

5. Put cutting plate in PC.

View attachment 4348877

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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Very nicely done. I was thinking about what happens when the PC cools ... If in non hepa air will it possibly suck in contamination?
 
Or_Gro

Or_Gro

Well-Known Member
led1k said:
Very nicely done. I was thinking about what happens when the PC cools ... If in non hepa air will it possibly suck in contamination?
Click to expand...
First, always let it cool naturally...never release pressure until opening.

Theoretically, the cooling process can suck in air....but I have found that my pot still has a little pressure at room temp, after overnight cooling...mine is made for this purpose, others may not hold a seal at room temp....

No harm, tho, letting it cool appropriately in your “cleanroom”...
 
Or_Gro

Or_Gro

Well-Known Member
Day 3: No evidence of contamination....yet.

C2EFC034-AB7A-450B-B5D2-CE407B1F7EC0.jpeg 81E60B5A-1743-41CB-9EF4-1DEC269495A2.jpeg D9FD10A6-ADE2-4D38-9CB9-B17FAB5B9058.jpeg

Running lights 24:0, @ 40-45 ppfd (1x96elite + 1x65w incandescent), to maintain steady heat @ ~75-77F, rh 50-60%...

5B42EF54-4B1C-425A-835C-ABD302204FFF.jpeg
 
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Or_Gro

Or_Gro

Well-Known Member
One of the common themes i’m reading in diff plant tissue culture (PTC) descriptions is use of 18:6 light schedules.

I’m used to running 24:0 during seedling/areocloning, and will be doing so here; so, i’ll find out if it makes a diff..
 
led1k

led1k

Well-Known Member
Or_Gro said:
One of the common themes i’m reading in diff plant tissue culture (PTC) descriptions is use of 18:6 light schedules.

I’m used to running 24:0 during seedling/areocloning, and will be doing so here; so, i’ll find out if it makes a diff..
Click to expand...
15:5 was mentioned somewhere... Maybe a faster day/night cycle brings them along faster?
 
Or_Gro

Or_Gro

Well-Known Member
Testing some small scale humidifiers, here’s first one:

Pulled stem apart and soaked tampon for 10 mins
76C59763-4D02-475B-A637-9042CC97CFF2.jpeg

Doesn't look like much
FB328F7F-C38E-41B7-95D4-9AF2B759D6CC.jpeg

Put it in this monster tote
3653C9E4-270E-4C96-84F0-630B5AAF4BB3.jpeg

Starting temp & rh
5CDFCD1B-6D26-493B-B096-338A742B89AC.jpeg

5 mins later
5C84E39F-9569-4F13-95FC-915342394EAA.jpeg 70FA4B3E-8FC8-4731-9B51-4CB2575885E6.jpeg

30 mins later
6FB5CCB5-597F-4CBE-B894-34A6615BF8AB.jpeg 42933067-C852-4F03-920C-026887881E4F.jpeg

Winner, winner, chicken dinner!

Manage rh level with a timer...

Here’s a 2’x3’x4.5’:
1786732E-56B2-445E-BE49-B7912BB5EC19.jpeg 9D97532F-D495-4D06-85D9-85D41722945D.jpeg
 
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Moabfighter

Moabfighter

Well-Known Member
Or_Gro said:
Cells are cells...
Click to expand...
Indeed. I’ve wondered this years ago I swear. Frickin cool to see someone trying to innovate it. Awesome job man. I’m actually in the market for a laminar flow hood, just trying to find one local ish for about 500 bucks......

Sending out a 2ml vial ready to plug in dirt whatever and go..... this IS what’s up man. Neat. Wow.
 
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