need colony formation assay protocol - (Jun/22/2010 )

Hi all,

Can anyone please send me protocol for colony formation assay ( not soft agar gel colony assay).
in this method colonies are stained with crystal violet or methylene blue after seedind in 6 well plate.
thank you..

YOu don't need a protocol to do this. After treatment, seed the cells at about 2000/well in 6-well plates and let the cell to grow for 10-12 days. Change medium every 2-3 days. At the end, stain the cells with crystal violet. The number of cells is critical--too many or too few is not good. Just do multiple repeats with varying number of cells/well.

Dear epigenetist,

thank you for help.
what I did is I seeded cells 1000 per well
next day added drug
after 48 hrs removed drug
changed medium.

Now I will do procedure as told by you. afer that how can I analyse my data? is there any formula to calculat percentage of colony formed in drug treated cells as compared to control?

i will appreciate your help.

you can count the number of colonies by eye or use some program such as imageJ after taking pictures.