This is for research and development, keeping these short, for preservation purposes
so they grow in petri dishes?
PTC, Organogenesis, or?
im not that advanced. I just grow and smoke and extract. I googled what you said. how do you observe that stuff? Lots of big microscopes or do you look at it on a computer screen attached. when I went to school we only had microscopes I at least know what a callus is but I dunno if seeds produce that.
It depends on what youāre trying to do. This should require a microscope, a very advanced jewelers scope, or one of the more advanced systems with a similar function
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Sure, Iāve done my fair share, but only with very old seed stock that wouldnt germinate on their own. Which usecase does @GenericWalrus need it for?
Somatic embryos generated from callus are way more efficient for multiplication and are even more similar to the donor than seeds as the only carry a slightly mutated version of the parents genome.
Epigenetics will also vary a bit due to generation pressure from tissue to callus to somatic embryo.
You could get one of those even to a homozyous state by creating a double haploid line using colchizin
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Also for all of this work, you need a really stable incubator with adjustable light / climate as they are very fragile
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@GenericWalrus is looking for tips on embryo rescue for old seeds.
If you havenāt already, you should probably hit the top of the threadā¦
5% sodium hypochlorite has been working fine, Iāll try 4.6% with hp.
== No contamination right?
But if it also == no growth, because youāre dealing with older material, can you be certain that being less aggressive wonāt help?
You are deliberately killing cells, with applied chemicals, with healthy fresh embryos you can provably kill a few cells and still have a viable embryo. At some point, killing even a few cells seems like it could tip the balance against you.
What do the fails look like? Is there a continuum?
Do you have a metric for success?
Are you scarifying?
According to the optimization process, 4.6% sodium hypochlorite along with 0.008% hydrogen peroxide for 16.81 min would result in the best outcomes. The results of a validation experiment demonstrated that this protocol resulted in 0% contamination as predicted, but germination rates were low and sporadic. However, using this sterilization protocol in combination with the scarification of in vitro cannabis seed (seed tip removal) resulted in 0% contamination and 100% seed germination within one week.
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I prefer to not utilize scarification unless absolutely necessary, with an abundance of seed stock available. I have seen some successes on the older seeds, but without question have seen the most failure from higher levels of aggression during scarification and sanitization. There have been more successes, to viable from seed from the in-vitro, with less agitation pre-germination or pre-multiplication, but this is likely from low trial levels in comparison. I would rather start with more seeds, use tried and true SOPās. For most newer seeds, this has not been an issue. For the 50-year-oldāseeds-plus, there is a much higher level of attention placed on the micro-nuances. Iām finding the viability is more pertinent to how the seeds were stored, before any type of attempt at germinating or micropropagation. I have 30 year old seeds from indoor, kept in vials with sanitation procedures and in darkness, that will germinate at 85%+, but 5 year old seeds from sun dried and market bought that have less than 15% successful germination rates, and when attempting to scarify these, they will disintegrate under pressure due to dryness regardless of the re-hydration techniques utilized.
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