bacterial mutation

[schwanda_lover]

hey guys! I am goign to be doign an experiment in the upcomign year relating to bactera and mutation. the basic premise is going to be testing whether bacteria control their rate of mutation depending on the amount of stress they are under. that is not the point of my post. I have most of the details of the experiment worked out, but one. I was thinking abotu it adn I don't know how to test for mutations. sin emutations occur on a DNA level, adn I probably won't have the equipment available to get to that level....what is a more general way to determine if the bacteria has undergone significant mutation....thanks.

[sdekivit]

you can isolate the DNA and do a restriction enzyme analysis (separation by gel electroforesis) and then do a Southern blot. You can then compare the fragments of good growing bacteria versus bacteria that are under stress.

[schwanda_lover]

ha - that seems so obvious...thank you.

[canalon]

Sdekivit give you one solution, here is another...

Measure the frequency of spontaneous resistant to antibiotics whose resistance can be acquired through mutation. Rifampicin is great. For detailed protocols, look for study by Matic, or Oliver and Baquero. This is a simple, method, quite reliable, although it involves quite a lot of plate counting

Good Luck

Patrick