Molecular identification of mixed powder of 9 kinds of Clematis medicinal materials based on high-throughput sequencing technology
Objective To establish a method for identifying 9 Clematis mixed powders based on high-throughput se-quencing technology.Methods The total DNA of 9 samples from Clematis was extracted and uesed ITS2 fragment for PCR amplification.The DNA fragments were separated by Illumina MiSeq platform.FLASH,QIIME,GraPhlAn and MEGA 7.0 software were used to sort and cluster DNA sequences.Results A total of 496 high-quality ITS2 sequences were obtained from the mixed powder samples,and 72 sequences were contained in the Clematis species.Seven species,including Clematis hexapetala,Clematis apiifolia,Clematis brevicaudata,Clematis fruticose,Clematis henryi,Clematis intricate,Clematis argenti-lucida,were compared,and Clematis manshurica and Clematis aethusifolia were not compared.Conclusion Using ITS2 as DNA barcode and high-throughput sequencing technology,seven species in the mixed samples of Clematis can be identified,which provides a basis for the identification of mixed medicinal materials.