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A double umbilical snail SNP molecular marker and its application in traceability

A technology of molecular markers and double-umbilical snails, which is applied in recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of small differences in allele distribution frequency, large heterozygosity, and high variability, and achieve The effect of rich polymorphism and small difference in frequency distribution

Active Publication Date: 2021-06-11
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] AFLP marker is the first generation of molecular marker method, which has rich polymorphism, strong genetic stability and high reliability, but it has high requirements on the quality and purity of DNA template and the level of operators, and it is difficult to count. The cost is relatively high; SSR marker is the second generation molecular marker method, which has the advantages of high polymorphism, low mutation rate and less template required, but the band pattern of the marker is complicated, the process is cumbersome, and it is not suitable for automatic typing ; and SNP, as a third-generation molecular marker, refers to the variation of a single nucleotide at the same site in the genome. It has the advantages of wide distribution, high density, strong representativeness, and good genetic stability. It does not require high template quality. Generally manifested as two alleles, showing either-or characteristics, easy to determine the type, and suitable for high-throughput automatic detection and typing, it is currently the most effective method for traceability molecular markers
[0005] However, the SNP molecular marker used for traceability is different from the general SNP molecular marker, and it must have at least the following characteristics: (1) high variability and close allele frequency; (2) small difference in allele distribution frequency between populations; (3) The degree of heterozygosity is greater than 0.3
Regarding traceable SNP molecular markers, patent CN201710607015.8 discloses a SNP marker combination and its application for traceability identification of beef cattle and meat products, and patents CN201510785854. SNP molecular markers used for traceability on chromosome No. 2 and their applications; patent CN201210529628.1 discloses a set of geographically specific Plasmodium vivax molecular markers and their application in strain traceability; Related reports on the SNP sites of snail traceability

Method used

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  • A double umbilical snail SNP molecular marker and its application in traceability
  • A double umbilical snail SNP molecular marker and its application in traceability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Search of SNP molecular markers

[0031] (1) Construction of DNA pool (pool)

[0032] Two umbilical snails in Shenzhen, Dongguan and Hong Kong were randomly collected respectively, and after the genomic DNA was extracted, the DNA of 6 individuals was aspirated and put into the same centrifuge tube, and mixed well for use.

[0033] (2) Primer design

[0034] Using the DNA sequence of the LOC106079843 gene of the umbilical snail (Genbank: NW_013487738.1) as a template, primers were designed to isolate the DNA fragment of the LOC106079843 gene of the umbilical snail. The primers are as follows:

[0035] Upstream primer: 5´-AAGAGTTGTGAGAGTGGGCC-3´ (as shown in SEQ ID NO.2)

[0036] Downstream primer: 5´-TCGGCCGAAAGTCATTATATGG-3´ (as shown in SEQ ID NO.3)

[0037] The total volume of the PCR reaction was 50 μL, including about 100 ng of A. umbilicus genomic DNA, 45 μL of 1.1×PCR mix, and 2 μL of 10 μmol / L upstream and downstream primers. The PCR amplification p...

Embodiment 2

[0044] Example 2 Distribution of alleles

[0045] (1) Design of test groups

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Abstract

The invention discloses a double umbilical snail SNP molecular marker and its application in traceability. The nucleotide sequence of the SNP molecular marker is shown in SEQ ID NO.1, which is 358 bp in total, and there is a base mutation 220C→220A at the 220th base, which is specifically recognized by the Fnu4HI restriction endonuclease. In 93 experimental populations of umbilical snails from 3 different locations, it was found that the allele frequency of the SNP molecular marker in different locations was close, the polymorphism was rich, the difference in allele frequency distribution was small, and the heterozygosity was greater than 0.3, it is preliminarily determined that the SNP molecular marker can be used for DNA traceability of the double umbilical snail.

Description

technical field [0001] The invention belongs to the technical field of molecular detection. More specifically, it relates to a double umbilical snail SNP molecular marker and its application in traceability. Background technique [0002] Double-umbilical snails belong to the phylum Molluscs, Pneumonia subclass, and Snail family. Among the more than 30 known species of double-umbilical snails, double-umbilical snails smooth, double-umbilical snails, shoal double-umbilical snails, and Puer's double-umbilical snails 18 species including S. mansoni and other 18 species can act as intermediate hosts of Schistosoma mansoni to spread schistosomiasis, causing serious disease burden to the world, especially in Africa, South America, Asia and other regions, and seriously hindering social and economic development. Among them, the double umbilical snails are mainly distributed in the fresh waters of St. Lucie in the Caribbean Sea and the northeastern coast of Brazil. They are highly su...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 吕志跃胡玥郑羽茜黄萍周洪利马玉斌周昱旻
Owner SUN YAT SEN UNIV
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