A pair of new universal 12S mitochondrial rRNA gene primers was designed through multiple alignment analysis of the mitochondrial tRNAPhe and the 5’ region of 16S mitochondrial rRNA genes of different kinds of fishes. The primers were successfully used to amplify an expected product fragment of about 1.2 kb from various marine fish species, and the amplified DNA fragment was recognized to contain the complete 12S mitochondrial rRNA and tRNAVal genes, as well as a partial 16S mitochondrial rRNA gene of about 146 bp in length. The primers would facilitate the study of the species discrimination, population and evolution in marine fish species.
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