disadvantages of soft agar assay - (Sep/14/2005 )
What are the possible disadvantages of soft agar assay other than the fact that the cells in the agar cannot be recovered?
Do you mean soft agar assay as a test for transformation? I don't know of disadvantages apart from that it needs to be performed very carefully to prevent fals-positives.
You can very easily isolate your cells from the soft agar. I did hundreds of clones in one day and they all survived and started growing in a dish.
How do you recover cells from sot agar?
Place your dish with soft agar under the microscope. Stick the pipettip of a p200 pipet into the agar and suck the clone into the pipettip.
Very easy to see the clone going into the pipettip[.
were you talking about soft agar in cloning, or soft agar for growing epithelial cell cultures?
I was referring to soft agar assay on cell lines to check for their transforming potential. I'm working with fibroblasts.
i tried picking out clones from the plate, but i could not get it to work. Once you have the clone in your pipette tip, then how do you proceed further. what kind of media do you use for these cells. Would they be the same for the ones you used for the original cells. I would really appreciate it if you can give me some pointers here.
hmmm...I have never done a soft agar assay for tissue culture cells; I am only experienced with soft-agar overlays with E coli. I regret that I am unable to help you
I was referring to theo who posted that he has picked hundreds of clones. i' m doing transformation assay for mammalian epithelial cells.
Aimikins, you said earlier that you have done this for fibroblasts. These are cell and tissue culture cells.
Aimikins i apologize for that slip up. I guess its SFHK doing fibroblast soft agar assay. once again, i apologize for that oversight.