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SNP molecular marker for biomphalaria and application of SNP molecular marker in traceability

A molecular marker, double umbilical snail technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragment, etc. Effects with small distribution differences and rich polymorphisms

Active Publication Date: 2019-02-15
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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  • SNP molecular marker for biomphalaria and application of SNP molecular marker in traceability
  • SNP molecular marker for biomphalaria and application of SNP molecular marker 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 an SNP molecular marker for biomphalaria and an application of the SNP molecular marker in traceability. The nucleotide sequence of the SNP molecular marker is shown in SEQ IDNO.1, is altogether 358 bp and has base mutation 220C or 220A at the 220th base, and the SNP molecular marker is specifically recognized by Fnu4HI restriction enzyme. In 93 biomphalaria test groups from 3 different locations, it is found that allele frequencies of samples of the SNP molecular marker in different locations are close, polymorphism is rich, difference in allele frequency distributionis small, and heterozygosity is larger than 0.3. It is preliminarily determined that the SNP molecular marker can be used for DNA traceability of the biomphalaria.

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 Applications(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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