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Sequencing analysis was used to characterize a mtDNA D-loop amplicon of 838 bp. Four pallid sturgeons, Scaphirhynchus albus, and four shovelnose sturgeons, S. platorynchus, from four states (Illinois, Louisiana, Montana, North Dakota) were used for this analysis. DNA sequencing of the nine fish revealed eleven polymorphic sites and six distinct haplotypes. No fixed nucleotide substitutions were observed between pallid and shovelnose sturgeons.