Bacterial Lipid Extraction via Bligh-and-Dyer

HPLC-grade organic solvents (Fisher Scientific, Ottawa, ON, Canada) and distilled deionized water were used throughout the experiment. Bacterial lipids were extracted following the Bligh-and-Dyer method^{15 (link)} with glass syringes, glass tubes, and ice-chilled solvents throughout the extraction process as previously described^{37 (link)}. After adding chloroform for monophasic extraction and eventual phase separation enhanced by centrifugation at 1,300 rpm for 5 min, the initial chloroform-rich phase was collected and supplemented with 0.5 mL 0.5 M sodium chloride (NaCl). After shaking by hand for 1 min, phase separation was again enhanced by centrifugation at 1,300 rpm for 5 min. The final chloroform-rich phase, approximately 4.2 mL, was collected for storage at −80 °C.