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Thread: Cheap Tissue Culture

  1. #1
    Sundrew's Avatar
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    Cheap Tissue Culture

    Hello TF,

    I'm pursuing a new cheap project! Recently, I was assigned a project in school where we have 7 weeks to pursue anything we want, and then we give a presentation on the process and what we have learned by the end. It's an awesome opportunity, and I have decided to move up my summer plans of attempting Tissue Culture. I am a student, and do not have access to any of the $15,000 laminar flow hoods or other equipment typically used. I'm not even going to try doing this with the typical MS agar and growth regulators. I'm going cheap, with a budget of $50. So, I'm going to have to get inventive and be careful, because I know how sensitive this practice is, and I by no means expect to be successful. But I'm going to try very hard to be, and hope I get it right the second time around . What follows is where I am in the process of acquiring materials.


    Today I bought most of the supplies I think I will need. These include:
    • A plastic box as a pseudo sterile hood as seen on the HTC webpage
    • 91% isopropyl alcohol
    • 70% isopropyl alcohol
    • Hydrogen peroxide (3% I believe)
    • 8.5% bleach (It was the best I could find, this was the super concentrated version too)
    • Inositol tablets
    • Multivitamin pills
    • Spray bottle to sterilize surfaces
    • Distilled water
    • Table sugar
    • Coffee filters to hold samples during sterilization



    As per very nice instructions from BigBella, I still need for my media:
    • Agar
    • 10:10:10 fertilizer



    In terms of equipment, all I need are:
    • Baby food jars
    • Long forceps
    • Pressure cooker



    For those that have taken the time to read this far, I really appreciate that. There are a few questions I still need answered. They are the following:
    • Are there alternatives to agar that I can also buy in a supermarket? I was informed that agar may not be available for a while around me. I could always order it if there is no alternative.
    • Is a 10% bleach solution 10% sodium hypochlorite or 10% of the bleach solution mixed with water (so would I just buy a 10% sodium hypochlorite version of Clorox Bleach, as opposed to mixing 10mL of the 8.5% Clorox Bleach I bought in 90mL of water)?
    • Is there any (preferably cheap) alternative to PPM I can use in the media to try and prevent unwanted growth? Any ideas with which I can experiment?



    Thank you so much for any help you can offer, and I will update this thread with the process and results.
    Last edited by Sundrew; 02-17-2013 at 08:21 PM.

  2. #2
    pokie22's Avatar
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    What equipment do you have access to? Balance, pressure cooker, pipets....?? Where are you located? I could send you some stuff to play with.

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    Sundrew's Avatar
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    I am fairly certain my school will let me borrow digital scales and graduated cylinders. I have a lot of pipettes, and I could probably get some nice glass graduated ones from school. I don't have access to things like an alcohol lamp or hot bead sterilizer. I'm located on the East Coast of the US. I'd be very grateful for anything you'd like to send me, as long as it is safe. I'm trying not to mess with the more powerful hormones and chemicals before I have a little experience with the cheap stuff.
    Last edited by Sundrew; 02-18-2013 at 08:06 AM.

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    31drew31's Avatar
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    Have fun with this project Drew. I bet the science department will have alcohol lamps, we had plenty at my high school.

  5. #5
    Sundrew's Avatar
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    Thanks Drew, I'm really excited for it. And if all goes well I should have a lot more of my brown D. burmanniis to spread around. I'll ask my science teachers if they have any lamps to spare.

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    Peatmoss's Avatar
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    Just make one yourself:

    <Av8tor1> as big as peat is, the bear runs not him

    Big Boss, Founder, and Major Cheese of the Canadian Association for the Cultivation of Carnivorous Plants... Ask if you want to join, I'm the only member...

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    Sundrew's Avatar
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    Cool Peat! What kind of alcohol do you use? Did you use anything special for the wick?

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    pokie22's Avatar
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    I can give you some gelrite, MS + vits powder, homemade PPM (use this judiciously as my results are not in but it is better than nothing), loops, small vials...etc. I do have baby food jars but they are heavy to ship, and personally I hate them. It is probably better to find a local free source than get them from me.

    As for the alcohol lamp, just use a lighter. You can dip/soak your instruments in alcohol and flame when necessary. You should have an adjacent bottle of sterile water to cool off the instrument before use.

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