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Metabolite extraction

LP Laura E. Peachey
CC Cecilia Castro
RM Rebecca A. Molena
TJ Timothy P. Jenkins
JG Julian L. Griffin
CC Cinzia Cantacessi
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Metabolites were extracted from 200 mg aliquots of each faecal sample using a methanol–chloroform–water (2:2:1) procedure, as described previously42. In particular, 600 μl of methanol–chloroform mix (2:1 v:v) were added, samples were homogenised using stainless steel beads and sonicated for 15 min at room temperature. 200 μl each of chloroform and water were added, samples were centrifuged and the separated aqueous and lipid phases were collected. The procedure was repeated twice, and the aqueous fraction from each extraction were pooled. The aqueous fraction was dried in a vacuum concentrator (Concentrator Plus, Eppendorf).

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