The invention discloses a method for amplifying mitochondrial
gene total sequences of fishes in Yangtze River based on a
degenerate primer combination and application thereof. The primer combination is obtained by designing in a conserved region through comparing the mitochondrial total sequences of fishes of acipenseridae, salangidae, cyprinidae, salmonidae, anguillidae, siluridae, bagridae, cyprinodontidae, hemiramphidae, synbranchidae, serranidae, gobiidae, channidae, bagridae and cobitidae, and the primer combination comprises Mt-1F, Mt-1R; Mt-2F, Mt-2R; Mt-3F, Mt-3R; Mt-4F, Mt-4R; Mt-5F,Mt-5R; Mt-6F, Mt-6R; and Mt-7F, Mt-7R. According to the method, the seven pairs of primers are used for carrying out PCR amplification on corresponding
gene fragments, and the mitochondrial
gene totalsequences are obtained through connection transformation, sequencing and splicing; the primer combination is less, the amplification effect is good, and the monitored fishes and roe can be effectively classified and identified. According to method provided by the invention, the degenerate primers are designed on the basis of the fish mitochondria conserved region, so that the mitochondrial gene total sequences of various fishes can be obtained; the universality is high, the obtained data can be applied to the research of fish mitochondrial genomes, thus perfecting a mitochondria
database.