Abstract
This protocol describes two similar methods for the extraction and methylation of fatty acids from bacterial cultures. The acid derivatization protocol (Lennen et al., 2013; Bligh and Dyer, 1959) results in the extraction and methylation of all fatty acids, both free and bound, from a bacterial culture, while the base derivatization protocol (Lennen and Pfleger, 2013) captures only bound (phospholipid, acyl-thioester) species. After extraction into hexane, the lipids may be analyzed by gas chromatography.
Keywords: Free Fatty Acid, Methylation, Fatty Acid Methyl Ester
Materials and Reagents
Equipment
Procedure
Notes
Acknowledgments
The protocol described here is based on the work of Bligh and Dyer (1959). The protocol was developed with funding from the US Department of Energy, Great Lakes Bioenergy Research Center.
References
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Authors indicated that they did not try to use that small volume of cell culture for this protocol, thus they did not have good answer to your question. Please share your experience with other readers via our "Reproducibility Feedback" if you successfully modify this protocol for that smaller vol. of cell culture in the future.Good luck,Bio-protocol team