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URL: https://dx.doi.org/DOI:10.17504/protocols.io.bc4qiyvw

Authors: Rowan Mclachlan, Agus Munoz-Garcia, Andrea Grottoli

Group: Coral Bleaching RCN protocols

Summary: This method is adapted and updated from methods originally published in Grottoli et al. (2004) and is based on the original methods of Folch & Stanley (1957), and Bligh & Dyer (1959). There are five parts to extracting lipids from ground corals: 1) grind and sub-sample the coral and store at -80 °C until ready to extract, 2) freeze-dry the sample, 3) extract the lipids from the freeze-dried samples, 4) standardize the lipid concentration to ash-free dry weight (AFDW), and 5) resuspend the extracted lipid for long-term storage and possible later analysis of lipid classes or isotopes. The lipid extraction procedure must be conducted in a fume hood with the sash as low as possible with the researcher wearing protective eyewear, gloves, and lab coat at all times.Important considerations regarding lipid analysis were gained from reading Chapter 1.3 “Lipid extraction, storage, and sample handling” from the textbook Lipid Analysis by Christie (2003). This method was originally developed by Andréa Grottoli and refined by Rowan McLachlan (06-11-18) with the guidance of Dr. Agus Muñoz-Garcia at The Ohio State University. This protocol was written by Rowan McLachlan (03-12-2020).ReferencesBligh, E. G., & Dyer, W. J. (1959). Canadian Journal of Biochemistry and Physiology. Canadian Journal of Biochemistry and Physiology, 37(8), 911–917. Christie, W. W. (2003). Lipid Analysis: Vol. 15, Isolation, Separation, Identification and Structural Analysis of Lipids. Current Opinion Biotechnology (3rd ed., Vol. 4). Oily Press Ltd, UK.Folch, J., Lees, M., & Stanley, G. H. S. (1957). A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry, 226, 497-509.Grottoli A. G., Rodrigues L. J., & Juarez C. (2004). Lipids and stable carbon isotopes in two species of Hawaiian corals, Porites compressa and Montipora verrucosa, following a bleaching event. Marine Biology, 145: 621–631.Christie, W. W. (2003). Lipid Analysis: Vol. 15, Isolation, Separation, Identification and Structural Analysis of Lipids. Current Opinion Biotechnology (3rd ed., Vol. 4). Oily Press Ltd, UK.Folch, J., Lees, M., & Stanley, G. H. S. (1957). A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry, 226, 497-509.Grottoli A. G., Rodrigues L. J., & Juarez C. (2004). Lipids and stable carbon isotopes in two species of Hawaiian corals, Porites compressa and Montipora verrucosa, following a bleaching event. Marine Biology, 145: 621–631.Folch, J., Lees, M., & Stanley, G. H. S. (1957). A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry, 226, 497-509.Grottoli A. G., Rodrigues L. J., & Juarez C. (2004). Lipids and stable carbon isotopes in two species of Hawaiian corals, Porites compressa and Montipora verrucosa, following a bleaching event. Marine Biology, 145: 621–631.

Affiliations: The Ohio State University, The Ohio State University, The Ohio State University

Version: 1

Publication Date: 2020

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