Phage amplificaiton


1. grow fresh log phase XL1 Blue cells by adding a single colony to 25 ml LB w/ 75 ul Tet, and grow in shaker until OD600 = 0.5-0.8 (~2x10^8 cells/ml).
2. add 10-100 ul phagemid (10^4-10^7 cfu) to 5 ml XL1 Blue.
3. incubate RT for 10 minutes.
4. add to 25 ml LB w/ 25 ul Carb and 75 ul Tet.
5. add ~10^10 helper phage (10-100 ul) moi~ 10-100:1
6. place in shaker for 30 minutes.
7. add 175 ul Kanamycin
8. Grow for 24-48 hours.
9. spin down cells @ 11K for 20 minutes and save supernatant. Titer phage.

note: OD600 = 100 for 3x10^10 cells/ml.


made Feb. 22, 1999; Baker Laboratory

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