Cauliflower cloning

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DoubleHelix

Hi all, prepping for cauliflower cloning. Followed the instruction for the media prep but cant help but feel it still feels quite gloopy and not set as well as I would expect it to be.
Has anyone else experienced this and have any advice?
 
DoubleHelix said:
Hi all, prepping for cauliflower cloning. Followed the instruction for the media prep but cant help but feel it still feels quite gloopy and not set as well as I would expect it to be.
Has anyone else experienced this and have any advice?
Click to expand...
It needs to be a 'soft set' so the students can push the cauliflower florets into the medium. I once made some with a bit more agar added and it was too firm which I thought would be ok but the students couldn't push the florets far enough into the medium.
If it is too soft then use less water or more agar.
 
Thanks all. Yes its a fine line between too firm that you cant get the explants into the medium and too runny they sink slightly!
Don't have too much experience with this experiment if i'm honest!
 
Using the SAPS method mentioned above worked we didn't have an issue with it setting.

Ensuring students insert the florets the correct way up helps and leaving vials where they can get light to aid growth seems to be important.
 
DoubleHelix said:
Hi all, prepping for cauliflower cloning. Followed the instruction for the media prep but cant help but feel it still feels quite gloopy and not set as well as I would expect it to be.
Has anyone else experienced this and have any advice?
Click to expand...
If the medium is too acidic (if the pH is too low, say below pH 5.0) then the agar can acid-hydrolyse during autoclaving, meaning that it will not set.

Ideally the pH needs to be about 5.8 before autoclaving.
 
The Half-Blood Prince said:
If the medium is too acidic (if the pH is too low, say below pH 5.0) then the agar can acid-hydrolyse during autoclaving, meaning that it will not set.

Ideally the pH needs to be about 5.8 before autoclaving.
Click to expand...
it isn't autoclaved, just microwaved for cauliflower cloning, then milton is added.
 
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