Method for identifying phytochemical type of marihuana by virtue of one-step multiplex PCR

A phytochemical type and cannabis technology, applied in the field of molecular biology, can solve problems such as the unseen cannabis chemical type, and achieve the effect of short cycle, low cost and simple operation

Inactive Publication Date: 2015-01-21
云南省农业科学院经济作物研究所
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  • Method for identifying phytochemical type of marihuana by virtue of one-step multiplex PCR
  • Method for identifying phytochemical type of marihuana by virtue of one-step multiplex PCR
  • Method for identifying phytochemical type of marihuana by virtue of one-step multiplex PCR

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Embodiment Construction

[0020] The technical solutions of the present invention will be further described below in conjunction with specific embodiments.

[0021] A method for identifying the chemotypes of marijuana resources from different sources in my country by a multiplex PCR reaction, comprising the following steps:

[0022] 1) Genomic DNA extraction of cannabis plants: The research materials are cannabis resources from different geographical sources in my country, which are planted in Kunming, Yunnan Province in the form of seed planting. The plant's conventional CTAB method is used to extract the cannabis genomic DNA, and the extraction object is the young leaf-like tissue of the cannabis plant in the rapid growth period. Take 0.5g leaf sample and grind it with liquid nitrogen or machine, and use the conventional 2×CTAB extraction method. After drying, the extracted DNA is dissolved in 12.5 μg / ml RNaseA aqueous solution, and placed in a water bath at 37°C for 1h. After the DNA solution was s...

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Abstract

The invention discloses a method for identifying a phytochemical type of a marihuana by virtue of one-step multiplex PCR. The method for identifying the phytochemical type of the marihuana by virtue of one-step multiplex PCR comprises the following steps: 1) extracting genome DNA of a marihuana plant; 2) designing a multiplex PCR primer; 3) preparing a PCR reaction system; 4) carrying out PCR amplification; 5) carrying out agarose electrophoresis; and 6) observing an electrophoretic band of a PCR product and judging a chemical type. By virtue of the one-step PCR reaction, a drug type marihuana can be amplified to obtain a specific THC stripe with the size being about 550bp, a fiber type marihuana can be amplified to obtain a CBD stripe with the size being about 1050bp, and an intermediate type marihuana can be amplified the THC stripe with the size being about 550bp and the CBD stripe with the size being about 1050bp at the same time, so that chemical type separation of a tested marihuana sample can be visually judged according to amplification stripes. The method for identifying the phytochemical type of the marihuana by virtue of the one-step multiplex PCR is not restricted by sampling time, a sampling site and the like in the traditional chemical detection, has the characteristics of short period, low cost and the like, is easy to operate, can be used for quickly identifying the phytochemical type of the marihuana and can be used by a public security drug detection department, an agriculture breeding department and the like, so that the method for identifying the phytochemical type of the marihuana by virtue of the primary multiplex PCR has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and relates to a method for identifying hemp phytochemical types by one-time compound PCR reaction. Background technique [0002] Cannabis (Cannabis sativa L.) is an annual herbaceous plant of the genus Cannabis (Cannabis) in the family Cannabinaceae, mostly dioecious. It is widely cultivated or wild all over the world, and now it is mainly distributed in Asia and Europe. my country's cannabis germplasm resources are rich, and it is a traditional economic crop in my country with a long history of cultivation. Hemp is a recognized double-sided plant. On the one hand, hemp has important economic value, involving textiles, papermaking, food, building materials, chemicals and pharmaceuticals, and has made great contributions to the development of human society. On the other hand, because the cannabis plant contains a hallucinogenic and addictive active ingredient - tetrahydrocannabinol (tet...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2537/143
Inventor 陈璇郭鸿彦杨明郭孟璧张庆滢许艳萍郭蓉陈裕
Owner 云南省农业科学院经济作物研究所
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