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in vitro culture clone
- Thread starter BonsaiBud
- Start date
wait....BonsaiBud said:
Imagine the possibilities. I suppose you need a healthy donor. I suppose it would need to be in veg.
what???
hows this stuff work?
Simple. You take advantage of the fact that plant cells are totipotent and can be treated as undifferentiated cells.
Create a sterile nutrient agar (murasaki & skoog, tweaked for cannabis).
Lace it with the correct hormone to coax those cells into becoming roots or shoots.
You will need to do BOTH, but since it is a different suite of hormones and required nutrients for each, this requires sterile transfer to a SECOND medium.
After you have tiny clusters of roots and shoots, called 'explants', they can be transferred and divided indefinitely.
TC begins with so little as a square cm of leaf that has been disinfected and pressed, bottom up, onto the medium. The stomata still must be able to breathe, as the tissue is still living and will be supported by the medium. It's primarily agar, and needs to be autoclaved before use.
But what do I know...
Here's the basic rundown:
http://www.omnisterra.com/botany/cp/slides/tc/tc.htm
Create a sterile nutrient agar (murasaki & skoog, tweaked for cannabis).
Lace it with the correct hormone to coax those cells into becoming roots or shoots.
You will need to do BOTH, but since it is a different suite of hormones and required nutrients for each, this requires sterile transfer to a SECOND medium.
After you have tiny clusters of roots and shoots, called 'explants', they can be transferred and divided indefinitely.
TC begins with so little as a square cm of leaf that has been disinfected and pressed, bottom up, onto the medium. The stomata still must be able to breathe, as the tissue is still living and will be supported by the medium. It's primarily agar, and needs to be autoclaved before use.
But what do I know...
Here's the basic rundown:
http://www.omnisterra.com/botany/cp/slides/tc/tc.htm
Wow. Just wow. Could you imagine how long you could keep genetics by using that method? Each clone would only need 2 inches of space in a lighted fridge. You could keep your favorite moms cloned in a safe place just in case something bad happens. Or you could keep cuts for future breeding purposes. Really interesting stuff.
H
humboldtlocal
Isn't that how they clone at the university of Mississippi. Or wherever that legal lab that grows medical marijuana licensed by the government is. I thought I saw that in one of those marijuana documentaries that was on tv last year.
I wonder if we could get ahold of their protocol for preparing their medium... hmmmm
GrowersUnite
Active member
pressure cooker?
M
medi-useA
...
muA
muA
What about a pressure cooker?
M
medi-useA
I have a new electric food cooker CROCK POT knockoff...Going to try that...and new fridge to put them in...now I gotta figure how to jerry-rig my fridge light to stay on or put a light in there...
muA
muA
I would put a flouro fixture in there. Just enough light to keep em happy and healthy. Not sure that your crock pot can reach the temps that are needed for sterilization, I know mine cant. Thats why people use the pressure cooker or autoclave.
M
medi-useA
Looks like I'll have to get a pressure cooker or an autoclave {enough buisnesses gone belly up so should be able to buy one @auction somewhere...}.
muA
Yea man I see them around from time to time. If you can, see if you can't find out how to get ahold of old college instruments because most of them are lightly used and well taken care of. I used to have a friend who picked up all kinds of stuff from colleges in his area. Everything you work with needs to be super sterile when doing this and make sure you take multiple cuts if its something you really want to save. Could be a valuable technique if you get it down pat. Hardest thing is getting the agar or whatever right with the right hormones to get it growing well. Let us know how it turns out.
M
medi-useA
will do...got some 'pecks paste' jars {glass w/metal lids}, agar, some other stuff, already...working on a Laminar Flow Hood and fridge/incubator @the moment...where are the alchohol lamp/torch thingies found?...and also anyone got ideas on where to get good metal tweezers from?
muA
muA
