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BL21 competent bacteria can be prepared by following the steps outlined below. Culture at 37@C until OD600 [?] 0. Spin out cells at 4@C Resuspend pellet in 15 mL ice-cold 0.1M MgCl2 Incubate on ice for 45 min.
BL21 competent bacteria can be prepared by following the steps outlined below. Culture at 37@C until OD600 [?] 0. Spin out cells at 4@C Resuspend pellet in 15 mL ice-cold 0.1M MgCl2 Incubate on ice for 45 min.
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BL21 competent bacteria can be prepared by following the steps outlined below. Culture at 37@C until OD600 [?] 0. Spin out cells at 4@C Resuspend pellet in 15 mL ice-cold 0.1M MgCl2 Incubate on ice for 45 min.
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1. Dilute an overnight culture of cells @ 1:100 in 50 mL of LB + chloramphenicol
2. Incubate shaking at 37˚C until OD600 ≈ 0.5 3. Spin out cells at 4˚C 4. Resuspend pellet in 15 mL ice-cold 0.1M MgCl2 5. Spin out cells at 4˚C 6. Resuspend pellet in 15 mL ice-cold 0.1M CaCl2 7. Incubate on ice for 45 min 8. Spin out cells at 4˚C 9. Resuspend pellet in 5 mL ice-cold 0.1M CaCl2, 30% glycerol 10. Aliquot into eppi tubes and snap-freeze 11. Store a –80˚C