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A method for detecting prickly ash with specific pcr primers

A specific, prickly ash technology, applied in the field of molecular biology detection of plants, can solve the problems of affecting the curative effect and the occurrence of adverse reactions, and achieve the effects of simple operation, high sensitivity and good specificity

Inactive Publication Date: 2018-07-24
重庆鑫跃丰农业开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Zanthoxylum bungeanum, bamboo leaf pepper, Evodia rutaecarpa, pepper, and mountain pepper are five different spice plants. If they are mixed or mixed into medicine, they will change their traditional medication habits, affect the curative effect, and even cause adverse reactions

Method used

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  • A method for detecting prickly ash with specific pcr primers
  • A method for detecting prickly ash with specific pcr primers
  • A method for detecting prickly ash with specific pcr primers

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1 is used for detecting the synthesis of the PCR primer of Zanthoxylum bungeanum

[0035] A total of 4 Zanthoxylum bungeanum ITS sequences were obtained from GenBank, and all Zanthoxylum bungeanum and green pepper samples were sequenced at the same time, and the above sequences were compared using the biological software DNAStar to find out the variation sites that distinguish Zanthoxylum bungeanum from other species. According to the position of the mutation site, primers of about 20 bases were designed on the upstream and downstream of the ITS sequence; the designed primers were analyzed with the biological software Primer 5.0, and the suitable primer pair for Zanthoxylum bungeanum was finally determined.

[0036] Zanthoxylum bungeanum specific primer pair:

[0037] (1) Upstream primer F1: 5'GCAATGCGAGCACTGCTT 3'

[0038] Downstream primer R1: 5'TCGTTCCCCGTGGGGGCGA 3'

[0039] (2) Upstream primer F2: 5'GCAATGCGAGCACTGCTT 3'

[0040] Downstream primer R2:...

Embodiment 2

[0050] Embodiment 2 Utilizes PCR primer to detect the specificity and sensitivity analysis of Chinese prickly ash

[0051] 1.1 Sample source

[0052] The medicinal material samples (see Table 1) used in this example were purchased from different pharmacies in Taiyuan City, Shanxi Province or from the Shanxi Academy of Traditional Chinese Medicine.

[0053] Table 1 Zanthoxylum bungeanum (green pepper) sample list

[0054]

[0055] 1.2 DNA extraction

[0056] Take about 60 mg of the above samples, grind them into powder with liquid nitrogen, and use the kit method to extract DNA. The kit was purchased from Beijing Aidelai Biotechnology Co., Ltd., model: CTAB Plant Genomic DNA Rapid Extraction Kit (Cat. No.: DN 14), 50 times.

[0057] 1.3 Serial dilution of sample DNA

[0058] The concentration of the above-extracted Zanthoxylum bungeanum DNA was detected to be 41.43ng / μl. The DNA samples were serially diluted, and the diluted concentrations were 4.143ng / μl, 2.0715ng / μl, ...

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Abstract

The invention provides a method for detecting zanthoxylum bungeanum maxim by using a specific PCR (Polymerase Chain Reaction) primer. Six pairs of specific primers which can detect the zanthoxylum bungeanum maxim quickly are designed on the ITS sequence of the zanthoxylum bungeanum maxim, a PCR detecting system is established on the basis of the six pairs of the specific primers, and about 520 bp single DNA (Deoxyribonucleic Acid) strips can be amplified, so that a molecular identification basis is provided for the herbal medicine identification of the zanthoxylum bungeanum maxim. The detecting method established according to the specific primers is easy to operate, high in specificity and sensitivity, and applicable to wide popularization and use, and can realize the detection of the zanthoxylum bungeanum maxim.

Description

technical field [0001] The invention relates to molecular biological detection technology of plants, in particular to a method for detecting Chinese prickly ash by using specific primer PCR. Background technique [0002] Zanthoxylum, a plant of the genus Zanthoxylum in the Rutaceae family, is a commonly used medicine for warming the interior in my country. . Zanthoxylum bungeanum has a long history whether it is used as medicine or food. It has been planted and used in China for more than 2,000 years. Zanthoxylum schinifolium Sieb.et Zucc. or Zanthoxylum bungeanum Maxim., which is recorded in the Pharmacopoeia of the People's Republic of China (2015 edition), is the dry and mature peel of green pepper Zanthoxylum schinifolium Sieb.et Zucc. or Zanthoxylum bungeanum Maxim. , abdominal pain caused by worms; external treatment of eczema, pruritus vulvae. Zanthoxylum bungeanum is a plant resource with high edible and medicinal value. It contains a variety of active ingredients ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/158
Inventor 张福生王丹丹许晓双秦雪梅张丽增
Owner 重庆鑫跃丰农业开发有限公司