A Study Of Fluorescent Multiplex Amplification Of 16srRNA And ND4 Genes In Mitochondrial DNA For Forensic Species Identification

Objective The purpose of our work was to establish a new fluorescent multiplex amplification of 16srRNA and ND4 genes in mitochondrial DNA, in order to provide a new method for forensic species identification. Methods The mitochondrial DNA sequences of human and different kinds of animals (include monkey, pig, dog, fish, rat, guinea pig, rabbit, horse, eel, frog, chicken, duck, goat, cow etc) could be obtained from GeneBank. By sequence-analysis software in GeneBank, the homology of mitochondrial DNA sequences between human and animals were analyzed. With primers design soft(Primer5), two pairs of primers were designed for 16srRNA and ND4 genes chosen from mtDNA sequence, and one of each pair of primers was marked by fluorescein(6-FAM) in 5'. Firstly, according to orthodox technique of multiplex amplification, a system of multiplex amplification was established, and then the amplifier products were analyzed by ABI PRISM 310 Genetic Analyzer, to establish...