Genetics - Preparation of Bacteriophage P1

Jeff Lawrence (G10)

1. Grow host strain overnight in LB at 37°.

2. Add 50 µL cells to 5 mL LB-Glucose:

	100 mL 	LB
	400 µL	50% Glucose

3. Incubate at 37° for 30 min with shaking.

4. Add 25 µL 1 M CaCl2 (5 mM Ca++ final concentration).

5. Add 100 µL P1-vir lysate titered at 5x108 pfu/mL, for a final concentration of 1x107 pfu/mL.

6. Incubate at 37° for 2 - 4 hours until culture clears.

7. Add 100 µL Chloroform and incubate at 37° for 2 min with shaking.

8. Centrifuge for 20 min at high speed at 4°.

9. Transfer supernatant to a screw cap tube. Add 50 µL Chloroform.

10. Store at 4°C. Titer drops after a few weeks.

Last Update: Thursday June 19 2014

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Eric Kofoid eckofoid at ucdavis.edu