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Specific PCR (Polymerase Chain Reaction) identification method of cordyceps militaris

A unique technology of Cordyceps militaris, which is applied in the field of authenticity identification of new resource health food, can solve the problems of restricting the industrial upgrading and standardized development of Cordyceps militaris, and difficulty in guaranteeing product quality, and achieves low cost, high accuracy, and strong primer specificity. Effect

Inactive Publication Date: 2014-02-26
CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is currently no standard method for authenticity identification and quality control of Cordyceps militaris, and it is difficult to guarantee product quality, which greatly limits the upgrading and standardized development of Cordyceps militaris industry

Method used

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  • Specific PCR (Polymerase Chain Reaction) identification method of cordyceps militaris
  • Specific PCR (Polymerase Chain Reaction) identification method of cordyceps militaris

Examples

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Embodiment 1

[0025] Example 1: Design of PCR primers and detection of samples

[0026] 1 Cordyceps militaris RPB1 Design of gene-specific PCR primers

[0027] According to the publicly registered Cordyceps militaris on GenBank RPB1 Gene sequence information for primer design (see attached figure 1 ).

[0028] The GenBank accession number of the reference sequence is: JN992491.

[0029] An upstream primer was designed at 13-32 and named as CICC-Cor-F1. The sequence is: 5'- AGCGAAAC CCGTCTATCACC -3'.

[0030] A downstream primer was designed at 410-429 and named as CICC-Cor-R1. The sequence is: 5'-CTTGGGGCCTTCTTCCGAAT-3'.

[0031]

[0032] 2 Detection of samples

[0033] 2.1 Collection of samples

[0034] Buy commercially available Cordyceps militaris, Cordyceps sinensis fungus powder, Paecilomyces bat moth powder, and Cordyceps sinensis.

[0035] 2.2 Sample genomic DNA extraction

[0036] Genomic DNA was extracted by the modified CTAB method and purified using the DNA purific...

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Abstract

The invention discloses a specific PCR (Polymerase Chain Reaction) identification method of cordyceps militaris. The technical scheme of the specific PCR identification method is as follows: genome DNA (Desoxyribo-Nucleic Acid) of a sample is taken as a template; a specific PCR primer designed by the specific PCR identification method is adopted to carry out one-time single-pipe specific PCR reaction so as to amplify the characteristic housekeeping gene of the cordyceps militaris; the actual amplification segment is 417bp; then the amplification product is detected through agarose gel electrophoresis to determine the authenticity of the sample. The specific PCR identification method is capable of specifically identifying the cordyceps militaris, and also capable of completing authenticity detection on the cordyceps militaris and the products thereof in different forms such as mycelium, fungus powder and capsules only through DNA extraction, PCR amplification and electrophoresis.

Description

technical field [0001] The invention belongs to the field of authentication of new resource health food, and in particular relates to an authentication method of Cordyceps militaris. Background technique [0002] The scientific name of Cordyceps militaris is Cordyceps militaris ( Cordyceps militaris ), is a famous and precious traditional Chinese medicine in our country. Its composition and efficacy are similar to those of Cordyceps sinensis. It can be processed into medicine and health products, and it is also a delicacy on the table, so it is widely loved by consumers. [0003] As a substitute for Cordyceps sinensis, Cordyceps militaris has the advantages of artificial cultivation, high yield and low cost, and has formed a large-scale production. According to statistics, the production of Cordyceps militaris in my country in 2012 was close to 8,000 tons, and the market size reached nearly 1.5 billion yuan. However, there is currently no standard method for authenticity i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2531/113
Inventor 程池扎西才吉李辉姚粟刘洋白飞荣
Owner CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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