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Thread: Cheap Tissue Culture

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    Sundrew's Avatar
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    So for PGRs I should probably buy the real deal. I guess those are chemicals I can't skimp on. What do fungicides do to the media/sample?

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    Quote Originally Posted by Sundrew View Post
    So for PGRs I should probably buy the real deal. I guess those are chemicals I can't skimp on. What do fungicides do to the media/sample?
    Just don't set yourself up to fail. There are a few things for which there are no real substitutes; and I would be willing to send you a couple of PGRs for your use -- an auxin and a cytokinin, say, 6-BAP and NAA, which are commonly used . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    Thank you very much for the offer. For this current project I think I will graciously decline, though it is much appreciated. I want to get the hang of lab procedure using inexpensive materials before I work with the actual chemicals, especially if they are someone else's.
    Last edited by Sundrew; 03-12-2013 at 03:24 AM.

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    I've been doing some research on cheap alternatives to synthetic PGRs. I could probably buy some young green coconuts from a local market. Coconut water is the liquid endosperm, rich in cytokinins, auxins, and other growth hormones. It also contains many carbohydrates that won't hurt the plants either. I found this paper that showed 10% - 20% coconut water resulted in greatly increased shoot formation in spinach. The problem with using coconut water is that there is not a specified volume to use, as it is extremely variable. Also, because I am not picking these coconuts off a tree for myself, the endosperm may have already solidified into the meat, or coconut milk. I'm going to look ridiculous walking around the market shaking coconuts...

    The study: http://www.academia.edu/1093292/Spin..._coconut_water

    Edit: Another study showing the exact compounds found in coconut water: www.mdpi.com/1420-3049/14/12/5144/pdf
    Last edited by Sundrew; 03-14-2013 at 09:17 AM.

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    Quote Originally Posted by Sundrew View Post
    I've been doing some research on cheap alternatives to synthetic PGRs. I could probably buy some young green coconuts from a local market. Coconut water is the liquid endosperm, rich in cytokinins, auxins, and other growth hormones. It also contains many carbohydrates that won't hurt the plants either. I found this paper that showed 10% - 20% coconut water resulted in greatly increased shoot formation in spinach. The problem with using coconut water is that there is not a specified volume to use, as it is extremely variable. Also, because I am not picking these coconuts off a tree for myself, the endosperm may have already solidified into the meat, or coconut milk. I'm going to look ridiculous walking around the market shaking coconuts...

    The study: http://www.academia.edu/1093292/Spin..._coconut_water

    Edit: Another study showing the exact compounds found in coconut water: www.mdpi.com/1420-3049/14/12/5144/pdf
    The issue with coconut water, is that it is, as you said, inconsistent; and the "exact" compounds vary more than many papers, or that table even suggests. Notice the variability of γ-Aminobutyric acid, for example, between a young green coconut (at 1.90 µg/mL and a mature one (at 1730.20 µg/mL) It can have stimulatory or inhibitory effects on some cultures; it can have greater auxin activity (through hydrolysis) if the media is autoclaved; lesser so, with zeatin and other compounds if the media is not sterile filtered instead. Many researchers avoid it due to that variability . . .
    Last edited by BigBella; 03-14-2013 at 05:12 PM.
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    I had no idea how I was going to measure out a volume, because there is no knowing what is actually in it. Plus, I checked three markets and there are no coconuts. Packaged coconut water has been pasteurized, so I doubt any of the cytokinins or auxins are still functioning.

    What do you mean by "it can have greater auxin activity (through hydrolysis) if the media is autoclaved?" I didn't really understand that.

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    Quote Originally Posted by Sundrew View Post
    I had no idea how I was going to measure out a volume, because there is no knowing what is actually in it. Plus, I checked three markets and there are no coconuts. Packaged coconut water has been pasteurized, so I doubt any of the cytokinins or auxins are still functioning.

    What do you mean by "it can have greater auxin activity (through hydrolysis) if the media is autoclaved?" I didn't really understand that.
    What I meant was that the active compounds in coconut water (which is generally considered an "undefined supplement" to many researchers) -- cytokinins, auxins, and the like -- are often chemically bound with other components in the coconut water; and that the process of hydrolysis, which occurs with autoclaving can "free-up" some of the available PGRs. Paradoxically, it can also serve to inactivate others.

    As an aside, the most significant component in coconut water is probably myo-inositol, which is readily available in any vitamin aisle . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    Myo-inositol, as in the Inositol I'm using for the original recipe you gave me? When I read about Inositol, there were few specific claims, but it appeared that it helped in nutrient uptake. Thank you for the explanation, that makes more sense. The chemicals hidden away in plants are astounding.

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