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Thread: Cheap Tissue Culture

  1. #17
    Sundrew's Avatar
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    Quote Originally Posted by BigBella View Post
    So, I assume that you will you have access to an autoclave or pressure cooker; and that that information on your blog, wasn't simply an illustration of the typical micropropagation process?

    Spores of microorganisms can survive very high temperatures, even sustained boiling, at normal atmospheric pressure. Generally, media has to be autoclaved at 121°C and 1 atm; or 15 psi; or 6.8 kg · 6.5 cm−2 for twenty-five minutes or more (depending upon the volume of the media) . . .
    Yes, I am in the process of finding a suitable pressure cooker, I should have one by this weekend. Sorry if this was not as detailed as your or pokie's great instructions, but I have to write it out in a way that my teachers and peers can understand. And thank you for all your help BigBella! Did you see your shout out?

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    Quote Originally Posted by Sundrew View Post
    Yes, I am in the process of finding a suitable pressure cooker, I should have one by this weekend. Sorry if this was not as detailed as your or pokie's great instructions, but I have to write it out in a way that my teachers and peers can understand. And thank you for all your help BigBella! Did you see your shout out?
    Great -- though ensure that the cooker can reach or exceed 15 psi; not all pressure cookers are capable of doing so. Good luck with the project and keep us posted . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

  3. #19
    Sundrew's Avatar
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    How long does it typically take to see initial growth on an explant? I won't expect seed germination, but over the course of a few weeks (3-5), can I expect to see some kind of growth? If, of course, there is no contamination.

  4. #20
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    Quote Originally Posted by Sundrew View Post
    How long does it typically take to see initial growth on an explant? I won't expect seed germination, but over the course of a few weeks (3-5), can I expect to see some kind of growth? If, of course, there is no contamination.
    It could take as little as three to four weeks; and that could be dependent upon the age of the leaf; how "savagely" it was sterilized; the temperature at which the culture are kept; and the concentration of the media, etc.

    Also, should the leaf experience phenolic bleeding (a darkening of the jello, not associated with contamination -- and that seems likely), keep moving it within the culture to a clear area, until that process stops . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

  5. #21
    Sundrew's Avatar
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    Thanks BigBella, that timing will be great.


    Here was the progress made today, I cut apart the plastic container to make my clean box. Here it is with the little spray bottle of bleach I will use to clean it and anything that enters it:

    DSC02115


    Here is the only 10-10-10 fertilizer I could find. It checked out from what I read on the back, I just don't know how to quantify this and add it to the gel mixture. Any suggestions?

    DSC02116

    DSC02118


    By next weekend I intend to have ordered some agar and have the liquid mixed. Hopefully I'll have the pressure cooker and forceps next week as well. Then I will have everything I need.
    Last edited by Sundrew; 03-03-2013 at 02:55 PM.

  6. #22
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    I ordered some proper fertilizer today, as well as some agar. I should be mixing the media by this weekend. In my research, I came across another potential experiment for a cheap gel, corn starch. It would create a firm and soft surface. The problem would be in spotting contamination on the cloudy surface, but here is a modified version that is clear: http://www.amazon.com/Barry-Farm-Cle...ords=clear+jel. I will probably be experimenting with normal cornstarch.

  7. #23
    Happy hour.. Api's Avatar
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    Good luck, Drew. Man, I would love to get my hands on a coffee flavored burmannii.
    Brennan
    My growlist: http://bit.ly/VUE0ri

  8. #24
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    Thanks. We'll see if it works...

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