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A kind of primer pair and application thereof for identifying giant centipede

A primer pair and centipede technology, applied in the field of molecular biology, can solve the problems of inability to adapt to the needs of the application, the length of time for the DNA barcode method, etc.

Active Publication Date: 2017-05-31
华润三九现代中药制药有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] With the development of molecular identification technology, DNA barcoding technology has been applied to PCR identification of spiny giant centipede and multi-thorny centipede (Zhang Hongyin, Chen Jun, Jia Jing, et al. Study on DNA barcoding identification of Chinese medicinal centipede and its counterfeit products, China Journal of Traditional Chinese Medicine, 2014,39(12):2208), but because the DNA barcoding method takes a long time and needs to be sequenced, it cannot meet the needs of the current on-site application of the molecular identification technology of traditional Chinese medicine
However, there has not been any report on the molecular specific identification between the less spiny giant centipede and the multi spiny centipede

Method used

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  • A kind of primer pair and application thereof for identifying giant centipede
  • A kind of primer pair and application thereof for identifying giant centipede

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Preparation and use method of a kit for identifying the giant centipede

[0048] 1. The design and synthesis of primer pairs used to identify the giant centipede

[0049] In this example, the universal primer COI (LCO1490: 5'-GGTCAACAAATCATAAAGATATTGG-3'; HCO2198: 5'-TAAACTTCAGGGTGACCAAAAAATCA-3') was selected to amplify and sequence the samples of the oligospinous giant centipede and the multispinous centipede, and then the sequence obtained was identical For source comparison, a pair of specific primers F1 and F2 are designed according to the variation region, which are used as specific primer pairs to identify the giant centipede. The sequence is as follows:

[0050] Primer F1: 5'-TCAATAATCGGAACAGCAT-3' (sequence 1);

[0051] Primer F2: 5'-ATACTGACCATACAAATAAAGGAG-3' (sequence 2).

[0052] 2. Assemble the kit used to identify the giant centipede

[0053] After the primers F1 and F2 designed and synthesized in step 1 are separately packaged, they are packaged in the...

Embodiment 2

[0066] Example 2. Using the kit prepared in Example 1 to identify the giant centipede

[0067] Samples to be tested: the two Chinese medicinal materials shown in Table 1, the giant centipede and the centipede.

[0068] 1. Extract genomic DNA from the sample to be tested

[0069] Take a sample of medicinal materials and disinfect the surface with 75% (volume fraction) ethanol. Use a sterilized scalpel to take 20 mg of the sample, then cut it with sterilized scissors, then use a grinder to grind and crush, and use marine animal genomic DNA The extraction kit [Tiangen Biochemical Technology (Beijing) Co., Ltd.] extracts total DNA. Please refer to the kit instructions for specific operations and store at -20°C.

[0070] 2. PCR amplification

[0071] Using the genomic DNA extracted from the sample to be tested in step one as a template, the primer pair (primer F1 and primer F2) designed in step one in Example 1 was used for PCR amplification. The specific reaction system and reaction condi...

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Abstract

The invention discloses a primer pair for identifying Scolopendra subspinipes multilans(L.)Koch and an application of the primer pair. The primer pair disclosed by the invention for identifying or assistant-identifying Scolopendra subspinipes multilans(L.)Koch is specifically the primer pair of two single-chain DNA molecules shown in sequences 1 and 2 in a sequence table. Experiments verify that by adopting the primers disclosed by the invention, Scolopendra subspinipes multilans(L.)Koch and Scolopendra multidens Newport are quickly and accurately identified by virtue of a quick PCR method and a fluorescent staining method and a fluorescent dye method is introduced to detect a true and false identification result, so that a technical support for field utilization on medicinal material molecular identification is realized.

Description

Technical field [0001] The invention belongs to the field of molecular biology, and relates to a primer pair for identifying the giant centipede and its application. Background technique [0002] According to a record in the Chinese Pharmacopoeia (2010), the origin of centipedes is the dried body of the centipede Scolopendra subspinipes mutilans L. Koch. Centipede is a valuable traditional Chinese medicinal material in my country. It has the effects of extinguishing wind, relieving spasm, dredging collaterals and relieving pain, attacking toxins and dispelling stagnation. Headaches, sores, scrofula, snake and insect bites, etc., mainly produced in Hunan, Hubei, Guangxi, Yunnan and other places in my country (Wang Keqin, Fang Hong, Ye Maoxiang, etc. Centipede drug source investigation and product identification, Chinese herbal medicine, 1997, 20(9):450). [0003] In recent years, centipede breeding has gradually emerged in my country. However, due to the uneven quality of most of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 黄璐琦袁媛崔占虎蒋超
Owner 华润三九现代中药制药有限公司
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