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A kind of Chinese Limulus ssr primer set and application thereof

A technology of primer sets and varieties, applied in the direction of recombinant DNA technology, biochemical equipment and methods, microbial determination/inspection, etc.

Active Publication Date: 2022-06-14
BEIBU GULF UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

There are relatively few technical methods for studying Chinese horseshoe crabs, and reports on the study of Chinese horseshoe crabs from the separation of microsatellite markers (Microsatellite DNA) are relatively rare. Due to the defects in the protection of Chinese horseshoe crab species, the environmental resources suitable for the growth of Chinese horseshoe crabs are declining day by day. Therefore, At present, there are very few samples used for the study of Chinese horseshoe crabs, and it is difficult to accurately reflect the true situation of the genetic diversity of Chinese horseshoe crabs; it also shows that the genetic diversity of Chinese horseshoe crabs is currently in its infancy, and these results cannot be used for the protection of Chinese horseshoe crabs. To provide a strong scientific basis, in order to conduct comprehensive and effective research on Chinese horseshoe crab, it is necessary to carry out the research work of this application, and to develop new research ideas for the SSR marker of Chinese horseshoe crab

Method used

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  • A kind of Chinese Limulus ssr primer set and application thereof
  • A kind of Chinese Limulus ssr primer set and application thereof
  • A kind of Chinese Limulus ssr primer set and application thereof

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Experimental program
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Embodiment

[0024] 1. Experimental method:

[0025] 1. Extraction method of Limulus genomic DNA:

[0026] Genomic DNA from Limulus blood and Limulus muscle were extracted by using Genomic DNA Extraction Kit from Blood Tissue (DP318-02) and Genomic DNA Extraction Kit from Marine Animal Tissue (DP324-03) produced by Tiangen Biochemical Technology (Beijing) Co., Ltd. , and diluted to 20ng·μL -1 , and stored in a -20°C refrigerator for later use.

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Abstract

The present invention relates to the technical field of molecular markers, in particular to a Chinese horseshoe crab SSR primer set and its application. The present invention extracts DNA from Chinese horseshoe crabs, and after sequencing and assembling, designs corresponding SSR primers to target multiple Chinese horseshoe crabs in the Beibu Gulf region. Genomic DNA was used as the material, and the designed SSR primers were verified, and 164 pairs of SSR primers were obtained that could amplify in Chinese horseshoe crabs. Among them, 20 pairs of polymorphic SSR primers were selected for molecular genetic markers of Chinese horseshoe crabs, which can distinguish multiple Chinese horseshoe crabs in the Beibu Gulf area. The gene of horseshoe crab sample, cluster analysis of different species of horseshoe crab in the Beibu Gulf area, provides new ideas for future research on genetic breeding, gene mapping, identification of species kinship and protection of genetic resources of horseshoe crab in China.

Description

【Technical field】 [0001] The invention relates to the technical field of molecular markers, in particular to a Chinese horseshoe crab SSR primer set and its application. 【Background technique】 [0002] Chinese Limulus has high economic and medical value. The Limulus reagent made from its blood can be used to detect bacterial endotoxins. Its meat, shell, and tail can be used as medicine. It is widely used in Guangxi, Hainan, Guangdong, Hong Kong, Fujian, and Taiwan in my country. , Zhejiang and other coastal provinces and regions are distributed. In recent years, due to human or non-human factors such as overfishing and environmental degradation, the number of horseshoe crabs in China has declined significantly, and they are on the verge of extinction. For a species, the higher its genetic diversity, the stronger its potential adaptability to changes in the living environment. The essence of Chinese horseshoe crab species protection is to protect its genetic diversity. Carryi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11C12Q1/6869G06K9/62
CPCC12Q1/6888C12Q1/6869C12Q2600/124C12Q2600/156G06F18/23C12Q2531/113
Inventor 王鹏良廖永岩许尤厚吴海萍朱鹏吴双成黄志兴苏洁霞李艺谢雅榆邓洁梅
Owner BEIBU GULF UNIV
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