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Wk5_Protocol - Protocol 1 Grow bacteria overnight in 10 mL...

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Protocol: 1. Grow bacteria overnight in 10 mL of the appropriate medium. (This will be done for you, but WHAT is the “appropriate” medium?). 2. Take 6 sterile micro tubes and add 1 mL of the bacteria into each tube and spin in the microfuge for 3 min. Remove the supernatant and dispose into empty beaker (save on ice). **Yes, you will have leftover bacteria; put it in the biohazard waste. 3. Resuspend pellet in 80 μL of 20% w/v sucrose, 0.03 M Tris-HCl pH 8.0. Add 20 μL of 5 mM sodium EDTA (pH 8.0). Mix by tapping on the bottom of the tubes and shake 10 min at room temperature. 4. Pellet cells in microfuge for 3 min. 5. Remove supernatant from each sample using a pipet, being careful not to disturb the pellet, and transfer to a SINGLE labeled micro tube. Store sample on ice. 6. Resuspend pellet in 100 μL of 50 mM sodium phosphate buffer and shake on ice for 10 min. 7. Pellet cells in microfuge for 3 min. 8. Transfer the supernatant of each sample into a second labeled micro tube.
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