Clonogenic Assay

How to perform a Clonogenic Assay.
Protocol by Peter Scarbrough: drive.google.com/file/d/15kq6...
Scarbrough PM, Mapuskar KA, Mattson DM, Gius D, Watson WH, Spitz DR. Simultaneous inhibition of glutathione- and thioredoxin-dependent metabolism is necessary to potentiate 17AAG-induced cancer cell killing via oxidative stress. Free Radic Biol Med. 2012 Jan 15;52(2):436-43. doi: 10.1016/j.freeradbiomed.2011.10.493. Epub 2011 Nov 4. PMID: 22100505; PMCID: PMC3664944.

[@mariaignacia650]

Hello! I'm doing a little research about clonogenic assay with colonies exposed to irradiation. If you could answer my question would be amazing. The colonies colored by the crystal violet, are the ones who are alive post irradiation? or am I wrong?

[@IowaRadOnc]

The colonies are stained using Coomassie blue. You wash off the media, wash with 70% ethanol for 5 mins and stain with the coomassie blue staining solution for 5 mins. The assay tests the cells ability to form clones (reproductive integrity). So yes the cells that survive form clones. You plot the surviving fraction ( normalized to your control group).