Protocol Online logo
Top : Forum Archives: : General Lab Techniques

Soft agar colony - Problems to stain (Sep/21/2008 )

Hello!

Do you make the working solution of crystal violet in water, methanol, ethanol? A final concentration of 0.005% is O.K.?
Thanks!

-Sergio-

I found the crystal violet concentration of 0.005% is too low and it takes a long time for cell to uptake it. If you prefer you can use a higher concentration. It is dissolved in water.

-pcrman-

QUOTE (pcrman @ Sep 21 2008, 05:10 PM)
I found the crystal violet concentration of 0.005% is too low and it takes a long time for cell to uptake it. If you prefer you can use a higher concentration. It is dissolved in water.

Thanks a lot... I will try a higher concentration, because I managed to get the experiment working once, but when I try to reproduce the experiment, I did not get any colony...

-Sergio-

You need to include a cancer cell line that is known to form colonies on soft agar as positive controls. Many things could affect colony formation such as type of agar, how agar is prepared (temperature, etc.), medium...

-pcrman-