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Thread: Starting Tissue Culture

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    Starting Tissue Culture

    So, mainly out of enjoyment, I'd like to try tissue culturing. I'd be most interested in propagating drosera and germinating nepenthes. I've done some research before, but is thereany recommended tutorial or supply source people could recommend? What pitfalls should I be on the lookout for?

    Thanks as always,

    Kyle

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    BigBella's Avatar
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    Quote Originally Posted by Vbkid View Post
    So, mainly out of enjoyment, I'd like to try tissue culturing. I'd be most interested in propagating drosera and germinating nepenthes. I've done some research before, but is thereany recommended tutorial or supply source people could recommend? What pitfalls should I be on the lookout for?

    Thanks as always,

    Kyle
    First and foremost -- and I am not joking -- have a fire extinguisher on hand, when flame sterilizing instruments and maintaining any open jars of 99% isopropyl alcohol. I keep one within arm's reach, mounted to the wall.

    Secondly, crack a book; and I wholeheartedly recommend Plants From Test Tubes, which is in its third or fourth edition by now. It was a standard of college horticulture classes. There are also myriad tutorials on YouTube; flytrapcare.com; on the ICPS site; but, first, learn those basics.

    Learn sterile technique; learn it. The biggest frustrations to novices are serial contamination issues, which can usually be attributed to ham-handed laboratory behavior. One clean room that I worked in had a chart on the wall outside, that described how plentiful and easily contaminants can be spread.

    Sitting or standing motionless, in the lab setting, allowed for 100,000 particles of 0.3 microns or larger to be sloughed off, per minute, in terms of regenerative things like skin cells, perspiration, and hair. Behavioral categories such as walking at 2.5 mph, produced 5,000,000 particles; 3.5 mph, 7,000,000; 5 mph, 10,000,000; and what had been described as "horseplay," produced 100,000,000 particles per minute; so no Greco-Roman wrestling during transfers.

    For the broader strokes, and for supplies, I would recommend hometissueculture.org. Their prices are generally far more reasonable than laboratory resources; and they now sell individual hormones in powdered form, along with KOH solutions, with which to dissolve the plant growth regulators, which are generally not water soluble. KOH -- potassium hydroxide -- is nasty stuff and is not called caustic potash without reason. Follow directions religiously.

    Invest in a decent pH meter; twenty bucks and a slow boat from Kowloon just won't cut it.

    Finally, determine where you will be doing your transfers -- whether it is a plastic tub or aquarium on its side; or whether you'll invest in building or buying a laminar flow hood. In the long run, you'll want a hood of some form . . .
    Last edited by BigBella; 05-28-2015 at 01:16 PM.
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    Chief Cat Behavior Specialist Knuckles's Avatar
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    The most satisfying feeling in the world is when you get your 1st batch to actually grow. You may have several that succumb to mold, bacteria, chemical overpowering, etc... Start slow & yes definitely grab a book as bella suggested. You can find cheap usb microscopes & build your own laminar flow hood w/supplies that you can find at your local hardware store. Find a nice spirit lamp or Bunsen burner, good scalpels, & forceps as well. You'll need a pressure cooker b/c we all don't have autoclaves & uv sterilizers lying around. Grab a bunch of baby food jars before you spend a bunch on containers. You may want to start with an easy species before you go chopping up your CPs. Nepenthes were especially hard for me when I first started out & I went back to good ol house plants like pothos & african violets. I began at age 13 & built my own mini lab in my bedroom closet. It was quite the accomplishment for a kid who had to fake his age & identity as a research lab owner on the phone with chemical companies to get my goodies. LOL. I started with murashige & skoog macro & micro nutrients, Basal salts, & a bunch of different anti-fungals, growth, & rooting formulas and way more others than what I needed. You do not need most of those at all & they need to be refrigerated away from foods.. These days there are premeasured mediums complete with everything you need for the type of plants you plan to tissue culture. These supplies were not as available to the public back then as they are today. All of this info + more is readily available online.

    Have fun & good luck.

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