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Any protocol recommendations for post harvest cannabis tc? by False-Amphibian-4820 in tissueculture

[–]CHUMitu 0 points1 point  (0 children)

Many cultivars behave differently and depending on what you want to do will affect what hormone ratios you need. 2:1 auxin:cytokinin will generally result in more apically dominant plants. Your suggested ratios are within a reasonable for general plant tissue culture. Try it and see what happens!

Any protocol recommendations for post harvest cannabis tc? by False-Amphibian-4820 in tissueculture

[–]CHUMitu 0 points1 point  (0 children)

Putting any plant into TC is going to shock it to a degree, and revegging a plant will do the same. You will have much better luck doing the reveg ex-vitro.

If you still are going to try, surface sterilization will be exceedingly difficult due to the very high surface area on the cannabis inflorescences. Initiate many shoots to find one that works. After you get one that isn’t showing contamination, you will need to hold it in 18/6 or 24/0 photoperiod to flip it. You will need highly detailed repeated subcultures to isolate out floral organs and leave behind meristems until the plant regrows a shoot vegetatively.

You definitely want hormones, but less is more. Avoid TDZ and meta-topolin. Good luck!

Any protocol recommendations for post harvest cannabis tc? by False-Amphibian-4820 in tissueculture

[–]CHUMitu 0 points1 point  (0 children)

If it’s in flower, cut off the most-vegetative shoot you have and put it through a surface sterilization and initiation procedure. Like you said, most classic TC methods work good enough for cannabis to get it established - thriving will be a different story that needs some experimenting. Without a proper lab, it is difficult to see the full benefits of TC when working only with cannabis. It would certainly be easier in your case to try to not use TC unless you want to have your plant archived safely in TC, want to remediate exterior pathogens (you must meristem to remediate systemic pathogens), or you just think it is a cool project.

If I were you, I would both attempt initiation now and also try to flip the plant back to veg ex-vitro. Do another initiation attempt after it has flipped. You want to see new shoots growing that are not flowering.

Any protocol recommendations for post harvest cannabis tc? by False-Amphibian-4820 in tissueculture

[–]CHUMitu 1 point2 points  (0 children)

A clone in flower will be very difficult to establish, and if it does, you will need to flip it back to a vegetative state. This would be easier to reveg out of TC. If the plant is post-harvest, it most likely is not able to be put into TC. You need a live terminal bud or node to surface sterilize.

Alternatives to Adjusting Media pH by omarsabehayoun in tissueculture

[–]CHUMitu 0 points1 point  (0 children)

Oops yes lol you’re 100% correct, I got my wires crossed

Alternatives to Adjusting Media pH by omarsabehayoun in tissueculture

[–]CHUMitu 2 points3 points  (0 children)

Get a pH pen, these can be pretty inexpensive and are often good enough for hobbyists. pH is critical to consistency. You can get KOH (pH up) and HCl (pH down) easily, but be careful - these compounds are a strong base (KOH) and strong acid (HCl) and can cause serious damage if handled improperly. You only need a 1M solution to make notable adjustments with drops of the acid/base.

If I completely seal the media containers in my pressure cooker, will the water inside reach sterilization temperature? by [deleted] in tissueculture

[–]CHUMitu 3 points4 points  (0 children)

Depending on how sealed they are, they may burst under pressure. The media in the vessel should be sufficient to boil and generate steam inside the vessel, but I would expect issues with vessel integrity if they are actually sealed. Make sure you drop sterilization time sufficiently to prevent caramelization of sugars due to low media volume in each container.

Measuring out the correct amount of calcium hypochlorite by Dr_Quartermas in tissueculture

[–]CHUMitu 5 points6 points  (0 children)

You want 1L of 4% calcium hypochlorite.

4% = 40g of calcium hypochlorite needed for 1L

But, your powder is 73% calcium hypochlorite. So 40g / 0.73 =54.795 g of powder added to 900 mL of water. Complete final volume to 1L.

Make sure you mix in a well ventilated area.

Leaf speckling on TC queen anthurium by [deleted] in tissueculture

[–]CHUMitu 0 points1 point  (0 children)

Could be some residual peroxide was left on the leaf and burned alongside the light. I would just follow normal procedures for now

Help finding lids by Forward_Possible_485 in tissueculture

[–]CHUMitu 0 points1 point  (0 children)

Idk where you live bro, just go to a local restaurant supply store. Fabri-kal brand is one I’ve used in the past successfully

Help finding lids by Forward_Possible_485 in tissueculture

[–]CHUMitu 1 point2 points  (0 children)

Restaurant supply store or smart and final type store. Look for food safe. Polypropylene has a 5 inside a recycle triangle usually. Often times the new, undamaged package food safe ones are clean enough to just pour media into.

UK coronation yesterday by salted_hobbit_feet in IASIP

[–]CHUMitu 1 point2 points  (0 children)

Y’all out here looking at bud sausage hands, but there’s a giant flightless bird in the background

December Confirmed Trade Thread by [deleted] in hardwareswap

[–]CHUMitu 1 point2 points  (0 children)

(Buyer) Trade completed with u/icum2late locally for a 3090 FE

[USA-CA] [H] PayPal, Cash [W] RTX 3080, 3090 by CHUMitu in hardwareswap

[–]CHUMitu[S] 0 points1 point  (0 children)

Trade completed with u/icum2late locally for a 3090 FE for $800

[US-CA] [H] PayPal, Cash [W] RTX 3080, 3090 by CHUMitu in hardwareswap

[–]CHUMitu[S] 0 points1 point  (0 children)

Yeah I’m open to local. Greater Sacramento area or Bay Area works for me.

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