Synergy Genetics - Tissue Culture Q & A

[SCJedi]

Here you go, I posted in-line, in BOLD:

@Rhizome

Many thanks SC_Jedi for the post and info!!!

I really would like to take this opportunity to ask you few questions and maybe enter into the topic.

• Benzlaminopurine (BAP) I Bought it…is it needed?

Not needed but recommended as an optional cytokinin!

• PPM I didn’t buy as it is very expensive ( BAP as well). I read some protocols are not using PPM.

If your aseptic technique is good, you dont need PPM or any biocides, period. It is expensive, can alter results in your experiments, and interact deleteriously with some PGRs.

• Protocols: any link to free sources? Protocols specific for different type of explants. leaves is more difficult that stems? Could be root?

Look at the links I pinned above. Most papers are NOT free but authors are often flattered if you email them and ask for a copy. There are a couple of meta studies out there that consolidate what a lot of research has done over the past 10+ years. One could aim for some of those but focus more on getting your chops down first. otherwise, you are going to have too many variables going at once

• Tricky one: what’s your opinion about mushroom incubator ( which simulate a laminar flow but without the HEPA filter, etc,etc,etc,etc) I like Plants in Jars YTVideos. + Kitchen well clean + pressur cooker as autoclave or microwave as steriliser.
  Which % of success would you believe I could achieve?

Again, only you can determine your level of success. I like Laur’s swagger, she picked how she wanted to do it and ran with it. If you want to use a SAB with a microwave and bleach in a kitchen garbage bag, go for it! The idea is to create a sterile environment. Do that with what ever level of funds you have available at the time or budgeted. Upgrades can happen any time but there is also a point of diminishing returns. TLDR; No need for an ISO rated lab here.

• Timing. After success can you control the grow to reduce or accelerate the passage to acclimation? Any link about timing from a leaf to be a 5’ cut. How long does it take? Can you use tissue culture and fridge to store genetics for long time ( 1-2 years before need to change medium)?

Yes, we all have lots of tricks up our sleeves but no, you won’t know how long a cultivar takes until you try it. For nodal it is typically 3 weeks for initiation, 3 for multiplication, and another 3-4 for rooting. If you are using meristem tissue expect it to take much longer but I am not sure why you would want something 5’ long in tissue culture. Did you mean inches and not feet?

Yes, you can use temperature as a tool to improve tissue culture goals in several ways. For example, often plants are chilled to simulate Fall/Winter and then forced into Spring-like conditions to get fast fresh growth. This method is referred to as temperature therapy and is often employed as an advanced technique (in conjunction with other steps) to eliminate pathogens from plant tissue

• DIY/Homemade: any link/info appreciated.

I will continue to add info but need to figure out what the top couple of blank posts will look like. I will likely wiki something and make a couple of repositories as well. However, here is a good one: Laminar Flow Hood - Build a HEPA filter flowhood | Fungifun

Many thanks!!!