The mitochondrion of the parasitic protozoonTrypanosoma bruceishows a number of unique features, many of which represent highly interesting research topics. Studies of these subjects require the purification of mitochondrial fractions. Here, we describe and discuss the two most commonly used methods to isolate mitochondria from insect stageT. brucei. In the first protocol, the cells are lysed under hypotonic conditions, and mitoplast vesicles are isolated on Percoll gradients; in the second method, lysis occurs isotonically by N2 cavitation, and the mitochondrial vesicles are isolated by Nycodenz gradient centrifugation.
