PCR specificity identifying primers and paris polyphylla identifying method adopting same

Abstract

The invention relates to a set of PCR specificity identifying primers and a paris polyphylla identifying method thereof and belongs to the technical field of molecular markers. The nucleotide sequences of the set of the PCR specificity identifying primers are CL-F: 5'- CGCGTACAACGAAACCATGCTGGGC-3' and CL-R: 5'- GAAGAGTGGGATGCCAACTGAGACC-3' respectively. The paris polyphylla identifying method adopting the PCR specificity identifying primers comprises the steps that 1, a genome DNA of a sample to be tested is extracted; 2, the genome DNA extracted in step 1 is used as a template and is subjected to PCR amplification by using the PCR specificity identifying primers; 3, the amplification product in the step 2 is subjected to electrophoresis detection, detect the bands at 130 bp for identification of the sample identified as dian floor, it is determined that the band to-be-identified sample detected out at the 130 bp position is paris polyphylla, and the band to-be-identified sample which is not detected out at the 130 bp position is other plant materials. The paris polyphylla identifying method only needs PCR, then gel electrophoresis is conducted on the PCR product, the paris polyphylla and other adulterants can be distinguished according to the size and existing situation of an electrophoretic band, and the method has the advantages of being simple and quick in operation and has unique advantages on the aspect of quick detection of medicinal material authenticity.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and in particular relates to a set of PCR-specific identification primers and a method for identifying the polyphylla. Background technique [0002] Paris L. is a perennial herbaceous plant of Liliaceae. According to the English edition of Flora of China, there are about 39 species of Paris L., which are mainly distributed in the tropical to temperate regions of Eurasia. The plants of this genus are mostly used medicinally in China, among which Parispolyphyllavar.yunnanensis(Franch.)Hand.-Mazz. and Huazhonglou[P.polyphyllavar.chinensis(Franch.)Hara.] were recorded in the 2005 edition "Chinese Pharmacopoeia", is one of the important raw materials of Yunnan Baiyao and other traditional Chinese medicines. [0003] However, the taxonomy of Pachyphysis plants is complex, because the characteristic characters of different groups overlap and overlap, and the boundaries of each type are blurre...

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Problems solved by technology

[0005] The above technology shows the possibility of using DNA barcodes in the identification of the authenticity of the medicinal materials of Yunnan Zhonglou. The present invention uses the technology of specific site PCR to overcome the problems of large sample size and high subjectivity in the traditional identification methods. There have not been any reports on molecular marker identification techniques for the identification of Chonglou, Changzhu Chonglou, Dali Chonglou, etc.

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