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Optical density measurement and isobutanol extraction

HX Hao Xie
PL Peter Lindblad
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Cell growth was monitored by measuring optical density at 750 nm (OD750) of each cell culture. The absorbance at 750 nm was measured every day for 200 μL cell culture in 96-well plate (SARSTEDT) using a microplate reader (HIDEX, Plate Chameleon).

Every second day, 2 mL of cell culture were sampled from each flask and centrifuged at 5000 rpm/2400×g for 10 min. Then, 1305 μL of supernatant were transferred into a 15 mL screw cap tube and mixed with 45 μL of 3000 mg L−1 internal standard 1-pentanol (Sigma-Aldrich) and 450 μL of extraction solvent dichloromethane (DCM, Sigma-Aldrich). The mixture was shaken on Multi-tube Vortexer VX-2500 (VWR) at maximal speed for 5 min, followed by centrifugation at 5000 rpm/4500×g, 4 °C, for 10 min. DCM phase was then transferred into 1.5 mL clear glass gas chromatography vials (VWR) for gas chromatography analysis.

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