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Week 08-Exp 7-Fall Break

Week 08-Exp 7-Fall Break - BIOL 519 Week 8 Generalized...

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BIOL 519 Week # 8- Generalized Transduction 1 BACKGROUND: On Tuesday of this week, you will first harvest the lysates you prepared last Thursday. Once you harvest the lysates, you will pass them through a 0.45 μ m filter. This filter has pores that are too small for bacteria to pass through, however, the pores are large enough for phage to pass through. This process will remove any viable CSH123 donor cells from your lysate, and leave only phage. You will then set up transduction reactions in which the P1 lysate will be added to a recipient strain (CSH108). The transduction reactions will be plated on Tetracycline plates to select for transductants that have gained the thiA ::Tn 10 from the donor strain, by way of the transducing particles in the lysate. Next week we will further test the transductants to determine whether they have also inherited the donor’s alleles at the rpoB and argE loci.
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BIOL 519 Week # 8- Generalized Transduction 2 TUESDAY Experiment 7 - Generalized Transduction with Phage P1 Harvest Lysates: Record the appearance of each of your lysate plates and the controls Collect the lysate from the plate that has completely lysed (if more than one has completely
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