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The pET3a containing hsEH CTD cDNA was transformed in chemically competent Ros2™(DE3) (EMD Millipore). A single colony of transformed cells was inoculated in 50 mL of ZYP5052 auto-induction medium [27] and cultured at 30 °C and 200 rpm overnight. |
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Protocol tips |
The pET3a containing hsEH CTD cDNA was transformed in chemically competent Ros2™(DE3) (EMD Millipore). A single colony of transformed cells was inoculated in 50 mL of ZYP5052 auto-induction medium [27] and cultured at 30 °C and 200 rpm overnight. |
Publication protocol
The pET3a containing hsEH CTD cDNA was transformed in chemically competent Ros2™(DE3) (EMD Millipore). A single colony of transformed cells was inoculated in 50 mL of ZYP5052 auto-induction medium [27] and cultured at 30 °C and 200 rpm overnight. The culture was diluted 1:100 in 3 × 1 L Erlenmeyer flasks filled up with fresh ZYP5052 medium and cultured at 37 °C and 200 rpm until the optical density of 0.5 was reached. The incubation temperature was then lowered to 18 °C and cells cultured for 24 h at 200 rpm. At the end of the incubation, the cells where centrifuged at 5000 × g for 30 min at 4 °C and the pellets stored at −20 °C.
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