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Found 1 matching solution for this experiment
pGEX-FIP-nha
Xuanming Tang, Institute of Agro-products Processing Science and
Upstream tips |
Protocol tips |
Downstream tips |
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The plasmid pGEX-FIP-nha was transformed into E. coli Rosetta (Novagen, CA, USA) competent cells. Transformants were grown at 25 °C for 6 h with shaking at 250 rpm in 200 mL of LB medium containing 0.1 mM IPTG and 50 µg/mL ampicillin. |
Cells were harvested by centrifugation at 5000× g for 5 min, suspended in 10 mM PBS (10 mL, pH 7.2), and disrupted by sonication. The crude extract was applied to a GST column (Pharmacia, Uppsala, Sweden), equilibrated and eluted with PBS to remove contaminants. |
Protocol tips |
The plasmid pGEX-FIP-nha was transformed into E. coli Rosetta (Novagen, CA, USA) competent cells. Transformants were grown at 25 °C for 6 h with shaking at 250 rpm in 200 mL of LB medium containing 0.1 mM IPTG and 50 µg/mL ampicillin. |
Downstream tips |
Cells were harvested by centrifugation at 5000× g for 5 min, suspended in 10 mM PBS (10 mL, pH 7.2), and disrupted by sonication. The crude extract was applied to a GST column (Pharmacia, Uppsala, Sweden), equilibrated and eluted with PBS to remove contaminants. |
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