Application of High-Throughput Sequencing Technology in Identification of Animal-Derived Species
Objective:High-throughput sequencing technology was used to establish an identification method of animal-derived species.Methods:Nine types of pure animal tissue samples were mixed with meat mince and DNA in proportion.The DNA samples extracted from the two mixed groups were labeled with barcodes and library preparation,followed by high-throughput sequencing and bioinformatics analysis.Finally,the combination with 1％DNA content was detected by real-time fluorescent PCR.Results:According to the Summary results,the sequencing results were good,9 kinds of animal origin mixed with 6 groups of meat paste were detected.Horses in DM 7-2 was not detected in 8 groups of DNA mixed samples,while other combined species were detected.And the proportion of reads in each species were basically as same as the proportion of initial DNA mixing.In the real-time fluorescence PCR experiment,the species with DNA ratio of 1％could be detected,and the CT value was between 32-35.Conclusion:A high-throughput sequencing method for identification of animal is preliminary established,with high accuracy and stability,and detection limit of 1％(mass fraction).
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