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Found 1 matching solution for this experiment
pET21b-TH2G
Arash Hatefi, Department of Pharmaceutics, Rutgers University
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To express biopolymers in BL21(DE3), BL21(DE3)pLysS or BL21(DE3) LOBSTR host, a single colony was picked and cultured in 5 mL Miller’s LB Broth (LB) starter culture containing 100μg/mL carbenicillin (Sigma-Aldrich Co. LLC., US). The starter culture tube was incubated overnight at 37°C under constant shaking at 350 rpm. The next morning, the whole starter culture volume was added to a flask containing 500 mL autoclaved Terrific Broth (TB) media (25.4g of TB powder, 2 mL of glycerol in 500 mL of Milli-Q water). The flask was shaken at 37°C/350 rpm and protein expression was induced at OD600 of 0.4–0.6 by 1mM Isopropyl β-D-1-thiogalactopyranoside (IPTG). |
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Protocol tips |
To express biopolymers in BL21(DE3), BL21(DE3)pLysS or BL21(DE3) LOBSTR host, a single colony was picked and cultured in 5 mL Miller’s LB Broth (LB) starter culture containing 100μg/mL carbenicillin (Sigma-Aldrich Co. LLC., US). The starter culture tube was incubated overnight at 37°C under constant shaking at 350 rpm. The next morning, the whole starter culture volume was added to a flask containing 500 mL autoclaved Terrific Broth (TB) media (25.4g of TB powder, 2 mL of glycerol in 500 mL of Milli-Q water). The flask was shaken at 37°C/350 rpm and protein expression was induced at OD600 of 0.4–0.6 by 1mM Isopropyl β-D-1-thiogalactopyranoside (IPTG). |
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