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Green silk turmeric PCR identification kit and identification method

A green silk turmeric and kit technology, applied in the field of DNA molecular identification, to achieve high sensitivity, short time-consuming and good repeatability

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

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Problems solved by technology

There is no report on the identification of green silk turmeric by PCR detection method

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  • Green silk turmeric PCR identification kit and identification method
  • Green silk turmeric PCR identification kit and identification method
  • Green silk turmeric PCR identification kit and identification method

Examples

Experimental program
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Effect test

Embodiment 1

[0029] 1. Acquisition of PCR primers for key reagents of green silk turmeric PCR detection method

[0030] Genomic DNA was extracted from Curcuma curcuma, Curcuma japonica, Curcuma curcuma Lvsi and Curcuma osmanthus with the method of rapid plant DNA extraction kit. Using the RAPD amplification method to perform RAPD analysis on the four types of turmeric, a specific amplified band of green silk turmeric was screened, and this specific amplification band only appeared in the green silk turmeric variety, and was not found in other turmeric varieties. There are no varieties, and the DNA molecule in this band is the specific DNA molecular marker of green silk turmeric. The green silk tulip specific DNA molecular marker was recovered from the agarose gel, cloned and sequenced. The sequencing result is shown in SEQ ID NO.1 and named LSY888.

[0031] According to the sequence of LSY888, design a pair of specific primers LSY1 and LSY2, the primer sequence is

[0032] LSY1:5'-CCTGAG...

Embodiment 2

[0046] 1. Study on the Accuracy of PCR Detection Method of Green Silk Turmeric

[0047] 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 performed using specific primers "LSY1 / LSY2". 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 green silk turmeric. The accuracy and reliability of PCR detection method, see the results figure 1 As shown, lanes 1 to 11 are Wen Yujin (Zhejiang Ruian), Wen Yujin (Zhejiang Yongjia), Huangsi Yujin (Sichuan Chongzhou), Huangsi Yujin (Sichuan Shuangliu), Green Silk Yujin,...

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Abstract

The invention discloses a PCR (polymerase chain reaction) identification kit and method for Radix Curcumae Phaeocaulis. Specific primers in the kit include LSY1 and LSY2, LSY1 is 5'-CCTGAGTGAGGACTTGTACG-3', and LSY2 is 5'-CGATGCTAAGGGCACAGCTA-3'. The invention provides the method of quickly and accurately identifying Radix Curcumae Phaeocaulis; the method is widely applicable to variety identification in the selection and breeding, planting, harvesting and storing, sales and clinical application of the Chinese herb, Radix Curcumae Phaeocaulis, and is significant in terms of Chinese medicine authentication, fake striking, Chinese medicine quality supervision reinforcement, and promotion of modern development of 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 the traditional Chinese medicine turmeric. [0003] (2) Background technology [0004] Green silk turmeric is the dried tuber of Curcuma pha eocaulis Val., and its authentic origin is Chongzhou, Sichuan. 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. Di...

Claims

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Application Information

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