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Guiyujin pcr identification kit and identification method

A kit, the technology of Guiyujin, applied in the field of DNA molecular identification, achieves the effect of short time consumption, strengthening the quality supervision of traditional Chinese medicine, and good repeatability

Active Publication Date: 2019-11-29
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Identification of Guiyujin by PCR detection method has not been reported yet

Method used

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  • Guiyujin pcr identification kit and identification method
  • Guiyujin pcr identification kit and identification method
  • Guiyujin pcr identification kit and identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Acquisition of PCR primers for key reagents of Guiyujin PCR detection method

[0030] Genomic DNA was extracted from Curcuma curcuma, Curcuma japonica, Curcuma curcuma Lvsi and Curcuma osmanthus with rapid plant DNA extraction kit. Using the RAPD amplification method to perform RAPD analysis on the four types of curcuma, a specific amplification band of Curcuma osmanthus was screened, and this specific amplification band only appeared in the species of Curcuma osmanthus. No, the DNA molecule in this band is the specific DNA molecular marker of turmeric. The specific DNA molecular marker of Guiyujin was recovered from the agarose gel, cloned and sequenced. The sequencing result is shown in SEQ ID NO.1 and named GYJ1176.

[0031] According to the sequence of GYJ1176, design a pair of specific primers GYJ1 and GYJ2, the primer sequence is:

[0032] GYJ1:5'-CTTCAACAACAGCACGACAA-3'

[0033] GYJ2:5'-GCTCAACAAACCCAAATACG-3'

[0034] The sites selected by the upstream an...

Embodiment 2

[0047] 1. Study on the Accuracy of Curcuma Curcuma PCR Detection Method

[0048] From warm turmeric (Ruian, Zhejiang), warm turmeric (Yongjia, Zhejiang), yellow silk turmeric (Chongzhou, Sichuan), yellow silk turmeric (Sichuan Shuangliu), green silk turmeric, osmanthus turmeric, ginger, Hainan notoginseng, grass fruit, Zhejiang Genomic DNA was extracted from Radix Paeoniae Alba and Atractylodes macrocephala plants. Using the genomic DNA of the above 11 materials as a template, PCR amplification was carried out using specific primers "GYJ1 / GYJ2". The PCR reaction solution, amplification procedure, and detection method were the same as in Example 1, except that the DNA template was replaced to verify the turmeric laurel PCR. The accuracy and reliability of the detection method, the results see figure 1 As shown, lanes 1 to 11 are Wen Yujin (Ruian, Zhejiang), Wen Yujin (Yongjia, Zhejiang), Yellow Silk Curcuma (Chongzhou, Sichuan), Yellow Silk Curcuma (Shuangliu, Sichuan), Green ...

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Abstract

The invention discloses a PCR (polymerase chain reaction) identification kit and a PCR identification method for radix curcumae kwangsiensis. Specific primers in the kit include GYJ1 and GYJ2, to be more specific, 5'-CTTCAACAACAGCACGACAA-3' and 5'-GGTTAAGCAATCTCTTCGGA-3' respectively. The method for rapidly and accurately identifying the radix curcumae kwangsiensis can be widely applied to varietyidentification in the steps of seed selection, breeding, planting, harvesting, storage, sales and clinical application, and has a great significance in identifying authenticity of traditional Chinesemedicine, combating counterfeits, enhancing quality monitor of the traditional Chinese medicine and promoting modernized development of the traditional Chinese medicine.

Description

[0001] (1) Technical field [0002] The invention relates to a DNA molecular identification method, in particular to a PCR identification kit, a specific primer, a template and an identification method of a traditional Chinese medicine turmeric laurel. [0003] (2) Background technology [0004] Curcuma kwangsiensis S.G.Lee et C.F.Liang is the dry tuber of Curcuma kwangsiensis S.G.Lee et C.F.Liang. Other varieties of turmeric include warm turmeric, yellow silk turmeric, and laurel turmeric. Curcuma is a traditional Chinese medicine, which has the effect of promoting blood circulation and removing blood stasis. It is used to treat amenorrhea, dysmenorrhea, chest and abdomen pain, jaundice, dark urine, epilepsy and madness. Modern studies have shown that turmeric can regulate immune function, anti-inflammation, inhibit the central nervous system, improve blood flow, etc., and can also be used to treat cancer, gastritis, and cardiovascular and cerebrovascular diseases. Different...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2565/125
Inventor 李敏洪露韩笑杰
Owner ZHEJIANG UNIV OF TECH
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