(r1) ProcedureBacterialMutationAccumulation < Lab

---+ 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

Barrick Lab > ProtocolList > ProcedureBacterialMutationAccumulation

Topic revision: r1 - 2012-01-21 - 15:40:56 - Main.JeffreyBarrick