Authentication of Dendrobium nobile by allele-specific diagnostic PCR / 中草药

ABSTRACT

Objective To develop a convenient and effective molecular authentication method for identifying Dendrobium nobile from other species of Dendrobium Sw.Methods Based on rDNA ITS sequences of D.nobile and other species of Dendrobium Sw.of Huangcaos and Fengdous,unusual sequence sites were confirmed and an allele-specific diagnostic primer pair was designed for D.nobile. Diagnostic PCR was performed using the primers with the total DNA of D.nobile and other species of Dendrobium Sw.as templates to establish the positive PCR condition for D.nobile.Results An about 300 bp DNA positive fragment was amplified from D.nobile with anneal temperature at 66 ℃ and anneal time at 40 s,whereas no any DNA fragment was amplified from the rest 39 species of Dendrobium Sw.Conclusion D.nobile could be effectively distinguished from other species of Dendrobium Sw.by diagnostic PCR with allele-specific diagnostic PCR,which is not only effective and specific,but also simple and timesaving,and has a comprehensive application prospects.