How to Perform Colony PCR
by
Ligations can be painful. Let’s just get that out of the way up front. When I was first learning them as a grad student they were significantly more painful because I had no idea how sensitive they were to the amount of DNA added. So as a beginner, I thought “I’m having trouble getting this insert to go in, I’ll just add more insert.” Bad call.
Not surprisingly, in my first year I performed many more “ligations” than “successful ligations.” With the number of minipreps I performed looking for that one colony that had my insert, I’m confident I singlehandedly propped up Qiagen’s quarterly revenue numbers for at least a year…
Then a brilliant new postdoc, Allart Stoop, joined the lab and taught me how to do molecular biology the right way. In addition to setting up the ligation reaction, Allart showed me the easiest way to analyze the results of the reaction….Colony PCR. Now, like a chain letter, we’re passing it on. Forward it to 10 friends in the next hour or your experiments won’t work for a year…
