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Tissue Culture success! (with seeds at least)

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Tissue Culture success! (with seeds at least)

Postby Matt » Sat Dec 20, 2008 4:40 pm

I wrote a bit of a journal of my first attempt at tissue culture. It's been a month now and some of the seeds are germinating, but unfortunately it doesn't look like any of the tissue will take. I'll know in another month probably. The blog might be a bit boring, but there are some photos at the end of it that everyone might looking at:
Tissue Culture success! (with seeds at least)
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Re: Tissue Culture success! (with seeds at least)

Postby Solarin » Sat Dec 20, 2008 5:00 pm

Stunning Matt,

This is really great news and I send a very heart felt "congratulations" with your success. Wow, what a Christmas present. Please keep us posted on the tissue culture results when they happen. If you need more seed, I have ~5K-8K or more in a glass vial from last year that I have in my fridge that I completely forgot about. Its yours if you need more material for further investigation. This really made my weekend guy.

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Re: Tissue Culture success! (with seeds at least)

Postby Matt » Sat Dec 20, 2008 5:05 pm

Thanks for the offer of the seeds Curtis! I have about 50 seeds right now that I'll probably sow the next time I do another attempt at culture, which will likely be in January. After that, I might send you a request for some more seed. Hang onto a couple hundred of those seeds for me.
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Re: Tissue Culture success! (with seeds at least)

Postby lemonlily » Sun Dec 21, 2008 2:16 am

Matt, congratulations!

Here comes the questions. Why do you have to do this?

For seeds:
1) 10% bleach for 15 minutes
2) 2% peroxide for 2 to 3 minutes
3) Rinse in sterile water
4) Transfer to media

For tissue:
1) Dip the cutting in the 90% alcohol quickly. Hold it above the alcohol to let any excess drain.
2) Drop the cutting in 3% peroxide for 2 minutes
3) Drop the cutting in 10% bleach mix for 15 minutes
4) Drop the cutting in sterile water for 10 minutes
5) Drop the cutting in 2% PPM (plant preservative mixture) solution for 60 minutes
6) Transfer to media

It sounds like you are killing it or something... I thought it was as simple as growing Dionaea in a container with seeds.
What is sterile? Why put it in bleach? What is peroxide?
Last edited by lemonlily on Sun Dec 21, 2008 2:49 am, edited 1 time in total.
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Re: Tissue Culture success! (with seeds at least)

Postby Matt » Sun Dec 21, 2008 2:45 am

I have to do the bleach and peroxide and PPM soaking to try to kill any bacteria or spores that are on the plant. If even one bacteria makes it into the culture jar, the culture is ruined. The bacteria just takes over and kills the seeds or plant material.

lemonlily wrote:What is sterile? Why put it in bleach? What is peroxide?

Sterile means free of bacteria and other micro-organisms. Bleach kills most bacteria, that's why I put the plant piece and seeds in bleach. Peroxide is hydrogen peroxide or H2O2. You can buy it at any pharmacy.
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Re: Tissue Culture success! (with seeds at least)

Postby linton » Sun Dec 21, 2008 3:10 am

Another triumph, well done Matt! It looks like all your hard work has paid off. I'm not sure if I would have the patience required for tissue culture. I might just have to stick with pullings and cuttings until I get them right. Good work and thanks for sharing your success with us.
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Re: Tissue Culture success! (with seeds at least)

Postby hackerberry » Sat Feb 14, 2009 5:41 am

Awesome Matt, keep up the good work!

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