DIY tissue culture

[Mega]

Hey.

I'd like to try to make a tissue culture.

According to this http://www.biologie.uni-hamburg.de/b-online/d29/29a.htm

The first tissue cultures were done using nutirent agar and coconut milk (for plant hormones).


So what I intend to do:
Mix LB-agar powder with coconut milk. Then autoclave. BUT: Do the phyto-hormones survive autoclaving?

Or should I just put it in the microwave and hope all bacteria are killed??

[Muralidharan E.M.]

 Two discussion groups that would be useful are the hometissueculture  http://tech.groups.yahoo.com/group/hometissueculture/  and the Plant-tc List Serve 

 

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[Muralidharan E.M.]

Sorry, I did not complete that.

Two discussion groups that would be useful are the hometissueculture group ( http://tech.groups.yahoo.com/group/hometissueculture/  ) and the Plant-tc List Serve at   http://plant-tc.cfans.umn.edu/listserv/listsrvh.htm 

You could do much better than  LB-agar powder + coconut milk by using some of the simple basal media and synthetic plant growth regulators like the cytokinins-Benzyl adenine and  Kinetin or auxins like IBA, 2,4-D. 

Yes, microwaving is a good DIY option but autoclaving is much better. Autoclaving will destroy only  some of the rarely used hormones and not the commonly used ones.

Murali

[Conner Berthold]

I would use a UV-C light source. I have a hood at home that uses a UV lamp to sterilize glassware and cultures. On the coconut cultures, I think that you should be fine with autoclaving, this article says that it should work. (Scroll to pg. 122) 

[Andreas Sturm]

Well Ok,


Surely I could buy some phytohormones. But aren't they expensive? I think rooting hormon solutions are avalable (cytokinin?) but where to get the other hormones??



I think the cheapest one is coconut milk....  and big advantage: all hormones etc. are included already (?).  So I can't forget any essential ingredient.

[Cory Tobin]

> Surely I could buy some phytohormones. But aren't they expensive? I think
> rooting hormon solutions are avalable (cytokinin?) but where to get the
> other hormones??

http://www.phytotechlab.com/searchresult.aspx?categoryid=10

I have purchased hormones through this company before. The prices are
reasonable even for small quantities (5g of auxin costs $14) and they
ship to residential addresses.


-cory