The Preliminary Study On DNA Barcoding Of Animal Chinese Traditional Medicine Earthworm

Objective:To extract genomic DNA from different sources Chinese traditional medicine earthworm in different methods,and compared these methods to determine which is the best method.Through PCR amplification,sequencing,sequence analysis and data analysis of different sources Chinese traditional medicine earthworm DNA Cytochrome c oxidase I gene(COI),16S rRNA gene(16S rDNA) sequences with universal primers, and construct phylogenetic tree,to provide reliable molecular markers of Chinese traditional medicine earthworm species identification and preliminary establishment of animal Chinese traditional medicine earthworm's DNA barcode.Methods:To Extract genomic DNA from different sources Chinese traditional medicines earthworm and select the most appropriate DNA exaction method of Chinese traditional medicine.Different sources Chinese traditional medicine earthworm DNA CO I,16S rDNA sequences with universal primers were PCR amplified and sequenced.Using DNAstar,DNAman,Geneious,Clustal X,MEGA softwares to analysis these DNA sequences, and construct phylogenetic tree.To submit the different sources Chinese traditional medicine earthworm DNA CO I,16S rDNA sequences to GenBank database DNA Barcoding database.Results:The suitable DNA extraction method of Chinese traditional medicine earthworm was determine.PCR amplification of different sources Chinese traditional medicine earthworm DNA CO I and 16S rDNA fragments was optimized,and obtain the different sources Chinese traditional medicine earthworm DNA COI,16S rDNA sequence and construct phylogenetic tree, different sources Chinese traditional medicine earthworm were clustered and analysised;then submit these sequences to GenBank database DNA Barcoding database.Conclusion:The obtained Chinese traditional medicine earthworm DNA COI,16S rDNA sequences can provide the molecular identification basis;the Chinese traditional medicine earthworm DNA sequence Co I polymorphism are more significant than the 16S rDNA sequence and are more suitable to be the Chinese traditional medicine earthworm DNA barcoding; to accumulated data.to the DNA barcode database of animal Chinese traditional medicine.