---+ Bacterial Mutation Accumulation Experiments ---++ Background Generally, you will need a large number of individual lines. The power of this experiment to measure mutation rates is the number of lines analyzed times the number of generations. ---++ Procedure ---+++ Setup Revive a culture of your strain by streaking it out on an agar plate and growing. Resuspend a single well-isolated colony in saline, dilute and plate so that you will have a new agar plate (or plates) containing several hundred well-separated colonies after another growth cycle. Also grow a liquid stock of this original culture to freeze. This is your original clone. ---+++ Daily transfers Transfer every day (or however many days every growth cycle takes). The timing should be as precisely the same as possible at eah transfer. Generally, stay within +/– 1 hr of the time of day from transfer to transfer. (*Very occasionally* you can leave the plates at 4°C for a day or two in case of emergency, but note than deviations from a well-controlled pattern, where the number of generations of growth, and their environmental conditions are kept constant at each transfer weakens the inferences that can be made from the data). ---+++ Archiving samples ---++ References
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Topic revision: r1 - 2012-01-21 - JeffreyBarrick