Due to its unique biology the mitochondrion of Trypanosoma brucei has attracted a lot of interest since many decades, making it arguably the best studied mitochondrion outside yeast and mammals. Here we describe a method allowing purification of mitochondria from procyclic trypanosomes that yields highly enriched and functional organelles. The method is based on isotonic lysis of cells by nitrogen cavitation, DNase I digestion, differential centrifugation and Nycodenz gradient centrifugation. The method is scalable and can be adapted to culture volumes a small as 100 mL or as large as 24 L.
CITATION STYLE
Niemann, M., & Schneider, A. (2020). A Scalable Purification Method for Mitochondria from Trypanosoma brucei. In Methods in Molecular Biology (Vol. 2116, pp. 611–626). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0294-2_36
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