Hi there...glad to hear it!! I'm told by my friends that do it on plants that the only tough part is getting them from the agar into the soil. I agree though, I would think that TC on plants would not be any harder than TC on human cells. It is just a matter of maintaining sterile technique which is not too difficult in truth. It will be interesting to see whether the same supplements will work for most plants....this is absolutely NOT true for mammalian cells of course. Have fun with it :o) Dan

Hi Klavier (Hi to you too Dan :) )

What are you using as your transfer cabinet?
I have looked at doing this too, and will no doubt be looking at again, but at the moment I am too busy with other stuff. The biggest initial expense seemed to be the purchase of a laminar flow cabinet and even construction of a suitable substitute wasn't cheap.
How do you intend doing your transfers so that you can maintain sterile conditions?

I recommend you get the book "plants from test tubes", and if you haven't seen these sites they might be of some help:

http://www.omnisterra.com/botany/cp/slides/tc/tc.htm

http://www.home.turbonet.com/kitchenculture/tcinfo.htm

Cheers, Troy.

What about the autoclave to sterilize your media and glassware? Are you using a pressure cooker? I'm not allowed to have a pressure cooker in the house because everyone KNOWS I would have way too much fun if the lid blew off....

What? I'm a walking pressure cooker! Use me, use me!

Werner - only amorphs? And what might fall into your "other large flowering plants" category?

Susan

For my cabinet, I am actually using the inside of my stove. I preheat the oven to 400 degress and then I let it back for 15-20 minutes with nothing inside. The inside is then entirely free of contamination and then I do all of my transfers inside of the stove while it is still very hot and I do it as quickly as I can. I actually fill the jars with medium days before I intend to use them, so I can see if any were contaminated. I place the jars in the stove and bake at 300 for 30 minutes. I place the lids in boiling water for the length of time it takes me to prepare the medium. I then, pour the boiling medium into the hot jars and seal with the lids strait from the boiling water. The jars tend to be so hot that the medium continues to boil for a few minutes even though it is away from the heat. When the jars cool, the air inside shrinks and vacume packs the jars. When I next open them, it is in the heated stove and with the sterilized plant material on hand. I do not touch anything (not just because most of the process is done at very high temperatures), and I use tongs and forcepts for everything. Just a note, the old mason jars with the glass lids do not work nearly as well as I thought they would. The Bell jars with the screw on lids are much better, and seal tighter. I do need to get smaller jars though, as the larger jars kind of waste medium and as I said it isn't cheap (at least not to me).

I forgot to mention, that I boil everything twice, occasionally, when I have more time three times (I do not have pressure cooker).

What I mean by big flowering plants is just about anything on Dan's website (except crinum lilies).

Hi Klavier,

Sounds exciting!! I'm up in Rochester now doing some expts on transgenic mice and have a bit of down time and so am goofing off!! I would think that if you asked in the Bio dept. they would have an autoclave you could use and that would make life easier. Also, any dept that does cell culture would have laminar flow hoods designed for doing TC and so if you ask nice they might let you use them or if you know the professor. That would make life way, way easier I would think, but you may not have access. Please let us know how the transfer to sterile soil goes as that wil be the exciting part. My interest in this is to be able to introduce compounds that could make 4n plants or perhaps use known agents that cause mutations in the plastid and therefore generate variegated material. Perhaps some day I can find the time to do this since I have all the materials necessary in my own lab...of course, then I couldn't fiddle around typing on GW as much :o) Dan

Still looking for plant material to play with.
Dan,
They do not let students use anything around here unless you are enrolled in a class that requires you to use it. If you come to them with something that to them seems like work and you don't want credit, they become very suspicious and the first thing they usually ask is if I am growing pot. What awful sterotyping of college students, and this is coming from the people that work with college students. You should have seen the look on there face when I carried 12 40Watt 6500K florescent bulbs into the dorm room with the new light hood I built. Two hours in the machine shop resulted in a florescent fixture that holds all twelve bulbs. I will have to post pictures of the homemade lab when I can. My next big project is making bio-diesel.

Hey Klavier,

Well, if you want to come to the U. of Pittsburgh and do a bit of TC in my lab you are welcome...just keep the Cannabis in NY :o) Dan

Thanks,
Kind of a far commute though. Would love to see your collection though (if I am ever in the area). I can only imagine where you put everything.

The professors are probably suspicious because the growing of cannibis is derived from an activity they participated in during their younger days. You know that people are most often suspicious of things they are doing or have a history of doing. You can't be suspicious of what you don't know about.

Susan