Freshwater bivalves in the superfamily Unionoidea possess distinct male (M)‐ and female (F)‐transmitted mitochondrial DNA (mtDNA). The former evolves independently of and at a significantly faster rate than the latter. Thus, population genetic and phylogenetic analyses of M sequences facilitate the generation of independent estimates of genetic variation and evolutionary relationships which are often more robust than those provided by analyses of F sequences alone. However, M mtDNA's rapid substitution rate often renders polymerase chain reaction (PCR) amplification difficult with ‘universal’ primers. Herein, we report on three pairs of PCR primers that consistently amplify the hypervariable M COII‐COI gene junction region in 25 bivalve genera (Unionoidea: Ambleminae).
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