FIG. 1.

B. mandrillaris multiplex rns PCR. Amplification was performed in 50-μl reaction mixtures with 2.5 mM MgCl₂; cycle conditions were 40 cycles of 1 min at 94°C, 2 min at 48°C, and 3 min at 72°C followed by a 15-min final extension at 72°C. Lanes 1, 6, and 10, 1-kb marker; lane 2, Balamuthia DNA with Balamuthia-specific primers; lane 3, Balamuthia DNA with Acanthamoeba-specific primers; lane 4, Acanthamoeba DNA with Balamuthia-specific primers; lane 5, Acanthamoeba DNA with Acanthamoeba-specific primers; lanes 7 to 9, Acanthamoeba and Balamuthia DNA with Balamuthia-specific primers (lane 7), with an Acanthamoeba-specific primer set (lane 8), and with both genus-specific primers sets (lane 9); lane 11, human DNA with both specific primer sets, lane 12, negative control. Amplicons produced by these mitochondrial 16S genus-specific primer sets are indicated on the right.