Week 08-Exp 7-Fall Break

1 biol 519 week 8 generalized transdu ction tuesday

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Unformatted text preview: ci. 1 BIOL 519 Week # 8- Generalized Transdu ction 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 lysed, use the one made with the most dilute phage) - Break up the top agar with a freshly flamed hockey stick - Using the hockey stick and a sterile spatula, transfer the top agar into a sterile 50 ml centrifuge tube - Wash the plate with 5 ml of sterile LB (spread it around with a freshly flamed hockey stick) - pour the LB into the same 50 ml centrifuge tube - TA will coordinate centrifugation of the lysates at 5,000 rpm for 10 min at 4oC - Filter sterilize the supernatant through a 0.45 µm filter (phage can pass through, but bacteria can not) into a sterile tes...
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