Figure 2. Multiplex polymerase chain reaction (PCR) for pheromone fragment analysis. A) Multiplex PCR with 4 sets of primers comprising MFα1 (V190/V191) and MFa1 (V290/V291, V1311/V1312, V1313/V1314) genes were carried out as described in Materials and Methods. Approximately 100-bp MFα1 and 117-bp MFa1 PCR amlicons were detected on 3.5% MetaPhor agarose in Tris-borate-EDTA buffer for MATα and MATa strains comprising Cryptococcus neoformans var. grubii (CnVG), Cryptococcus neoformans var. neoformans (CnVN), and Cryptococcus gattii (Cg). Lanes 1 and 7, molecular mass marker. B) Multiplex PCR depicting MFα1 and MFa1 PCR amplicons from the 8 known hybrid (A/D) isolates. Lane 1, molecular mass marker.