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Analysis of polar lipids

Polar lipids are extracted from freeze dried cell material using a choroform:methanol:0.3% aqueous NaCl mixture, polar lipids are recovered into the chloroform phase (modified after Bligh and Dyer, 1959).
Polar lipids are separated by two dimensional silica gel thin layer chromatography. The first direction is developed in chloroform:methanol:water, and the second in chloroform:methanol:acetic acid:water. Total lipid material is detected using molybdatophosphoric acid and specific functional groups detected using spray reagents specific for defined functional groups (Tindall et al., 2007). The labelled picture contributes the report.

Required material: For this method 200 mg freeze dried cells are needed or 1 g wet biomass send in frozen state on dry ice or suspended in isopropanol (send at ambient temperature). Alternatively, an actively growing culture (still in exponential growth phase upon arrival, without signs of sporulation or cell lysis) could be send. Subculturing (if required) will be charged separately. If subculturing is requested by the customer, detailed cultivation conditions (media, temperature, oxygen supply, cultivation duration) must be provided.

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  1. Bligh, E.G. and Dyer, W.J. (1959). A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol., 37, 911-917
  2. Tindall, B.J., Sikorski, J., Smibert ,R.M., and Kreig, N.R. (2007) Phenotypic characterization and the principles of comparative systematics. In Methods for General and Molecular Microbiology 3rd edn. Pp. 330-393.